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US20180312473A1 - Phenazine derivatives as antimicrobial agents - Google Patents

Phenazine derivatives as antimicrobial agents Download PDF

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Publication number
US20180312473A1
US20180312473A1 US15/744,319 US201615744319A US2018312473A1 US 20180312473 A1 US20180312473 A1 US 20180312473A1 US 201615744319 A US201615744319 A US 201615744319A US 2018312473 A1 US2018312473 A1 US 2018312473A1
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alkyl
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Robert William Huigens, III
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University of Florida Research Foundation Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/36—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems
    • C07D241/38—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems with only hydrogen or carbon atoms directly attached to the ring nitrogen atoms
    • C07D241/46—Phenazines
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/48—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with two nitrogen atoms as the only ring hetero atoms
    • A01N43/60—1,4-Diazines; Hydrogenated 1,4-diazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/498—Pyrazines or piperazines ortho- and peri-condensed with carbocyclic ring systems, e.g. quinoxaline, phenazine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/005—Antimicrobial preparations
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/36—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/36—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems
    • C07D241/38—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems with only hydrogen or carbon atoms directly attached to the ring nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/36—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems
    • C07D241/38—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings condensed with carbocyclic rings or ring systems with only hydrogen or carbon atoms directly attached to the ring nitrogen atoms
    • C07D241/40—Benzopyrazines
    • C07D241/42—Benzopyrazines with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring

Definitions

  • biofilms In addition to infections resulting from planktonic bacteria, biofilms also play a key role in pathogenesis.
  • the NIH has stated that bacterial biofilms are associated with up to 80% of all bacterial infections. Biofilms are notorious for their resistance to conventional antibiotic treatments.
  • innovative antimicrobial strategies are needed to meet the biomedical challenges of microbial infections, especially those resulting from multidrug resistant microbial infections and pathogenic bacterial biofilms.
  • the present invention provides novel halogenated phenazine derivatives (HPs, HP analogues), such as compounds of Formulae (I′) (e.g., Formula (I)), (II), and (III), and salts, hydrates, solvates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, prodrugs, thereof:
  • Exemplary compounds of the invention include, but are not limited to:
  • Exemplary compounds of the invention also include, but are not limited to:
  • Exemplary compounds of the invention also include, but are not limited to:
  • Exemplary compounds of the invention also include, but are not limited to:
  • Exemplary compounds of the invention also include, but are not limited to:
  • the compounds of the invention may exhibit antimicrobial activity (e.g., antibacterial activity, such as antibacterial activity against strains of Staphylococcus aureus (e.g., methicillin-resistant strains of Staphylococcus aureus ), strains of Staphylococcus epidermidis (e.g., a methicillin-resistant strain of Staphylococcus epidermidis (MRSE)), and strains of Enterococcus faecium (e.g., vancomycin-resistant strains of Enterococcus faecium )).
  • antimicrobial activity e.g., antibacterial activity, such as antibacterial activity against strains of Staphylococcus aureus (e.g., methicillin-resistant strains of Staphylococcus aureus ), strains of Staphylococcus epidermidis (e.g., a methicillin-resistant strain of Staphylococcus epidermidis (MRSE)
  • the compounds of the invention may act by a microbial warfare strategy (e.g., a reactive oxygen species (ROS)-based competition strategy) similar to the one employed by Pseudomonas aeruginosa ( P. aeruginosa ).
  • the inventive compounds may generate ROS in, near, or around a microorganism (e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite), which may be toxic to the microorganism.
  • a microorganism e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite
  • inventive compounds may be able to reduce, inhibit, and/or remove biofilms (e.g., Staphylococcus aureus biofilms (e.g., MRSA biofilms) and/or Staphylococcus epidermidis biofilms (e.g., MRSE biofilms)).
  • biofilms e.g., Staphylococcus aureus biofilms (e.g., MRSA biofilms) and/or Staphylococcus epidermidis biofilms (e.g., MRSE biofilms
  • the inventive compounds preferably have minimal or no adverse side effects.
  • the inventive compounds have low cytotoxicity with respect to mammalian cells and/or demonstrate low hemolysis activity.
  • the present invention provides compositions including a compound of the invention and optionally an excipient.
  • the composition includes an effective amount of the compound for disinfecting a surface.
  • the composition is a pharmaceutical composition including a compound of the invention and optionally a pharmaceutically acceptable excipient.
  • a pharmaceutical composition of the invention includes an effective amount of a compound of the invention for administration to a subject.
  • the pharmaceutical composition is useful in a method of the invention (e.g., a method of treating a microbial infection, preventing a microbial infection, inhibiting the growth of a microorganism, inhibiting the reproduction of a microorganism, killing a microorganism, inhibiting the formation and/or growth of a biofilm, reducing or removing a biofilm, or disinfecting a surface).
  • the microorganism is a microorganism described herein.
  • the microorganism is a bacterium.
  • the bacterium is a Gram-positive bacterium (e.g., a Staphylococcus species or Enterococcus species).
  • the bacterium is a Gram-negative bacterium (e.g., an Acinetobacter species).
  • the microorganism is a mycobacterium (e.g., a strain of Mycobacterium tuberculosis ).
  • Another aspect of the present invention relates to methods of treating and/or preventing a microbial infection in a subject in need thereof, the method including administering to the subject a therapeutically or prophylactically effective amount of a compound or pharmaceutical composition of the invention.
  • the microbial infection is treated and/or prevented by the inventive methods.
  • the microbial infections that may be treated and/or prevented by the inventive methods include, but are not limited to, microbial respiratory tract infections, microbial gastrointestinal tract infections, microbial urogenital tract infections, microbial bloodstream infections, microbial ear infections, microbial skin infections, microbial oral infections, microbial dental infections, microbial wound or surgical site infections, microbial infections associated with cystic fibrosis, and microbial infections associated with implanted devices.
  • the microbial infection described herein is a bacterial infection.
  • the bacterium causing the bacterial infections is a Gram-positive bacterium (e.g., a Staphylococcus species or Enterococcus species).
  • the bacterium causing the bacterial infections is a Gram-negative bacterium (e.g., an Acinetobacter species).
  • the microbial infection described herein is a mycobacterial infection (e.g., an infection caused by Mycobacterium tuberculosis ).
  • the subject is a human.
  • the subject is a human with cystic fibrosis.
  • the subject is a non-human animal.
  • the present invention provides methods of inhibiting the growth of a microorganism (e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite) in vitro or in vivo.
  • a microorganism e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite
  • the present invention provides methods of inhibiting the reproduction of a microorganism (e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite) in vitro or in vivo.
  • a microorganism e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite
  • the present invention provides methods of killing a microorganism (e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite) in intro or in vivo.
  • a microorganism e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite
  • an inventive method includes contacting a microorganism (e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite) with a compound or pharmaceutical composition of the invention in an amount effective at inhibiting the growth and/or reproduction of or killing the microorganism.
  • a microorganism e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite
  • biofilm includes a microorganism (e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite).
  • the biofilm includes bacteria.
  • the biofilm may include one or more species of bacteria and/or other microorganisms.
  • Another aspect of the present invention relates to methods of disinfecting a surface, the methods including contacting the surface with an effective amount of a compound or composition of the invention.
  • the surface is a biological surface (e.g., skin).
  • the surface is a non-biological surface.
  • kits comprising a container with a compound or composition (e.g., pharmaceutical composition) of the invention.
  • the kits of the invention may include a single dose or multiple doses of the compound or pharmaceutical composition thereof.
  • the provided kits may be useful in a method of the invention (e.g., a method of treating a microbial infection, preventing a microbial infection, inhibiting the growth of a microorganism (e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite), inhibiting the reproduction of a microorganism, killing a microorganism, inhibiting the formation and/or growth of a biofilm, reducing or removing a biofilm, or disinfecting a surface).
  • a kit of the invention may further include instructions for using the kit (e.g., instructions for using the compound or composition (e.g., pharmaceutical composition) included in the kit).
  • the present invention provides uses of the compounds and pharmaceutical compositions of the invention for manufacturing a medicament for treating and/or preventing a microbial infection.
  • the present invention provides the compounds and pharmaceutical compositions of the invention for use in methods of preventing and/or treating a microbial infection.
  • the present invention provides the compounds and pharmaceutical compositions of the invention for treating and/or preventing a microbial infection.
  • Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, e.g., enantiomers and/or diastereomers.
  • the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer.
  • Isomers can be isolated from mixtures by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses.
  • HPLC high pressure liquid chromatography
  • C 1-6 is intended to encompass, C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 1-6 , C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-5 , C 2-4 , C 2-3 , C 3-6 , C 3-5 , C 3-4 , C 4-6 , C 4-5 , and C 5-6 .
  • aliphatic refers to alkyl, alkenyl, alkynyl, and carbocyclic groups.
  • heteroaliphatic refers to heteroalkyl, heteroalkenyl, heteroalkynyl, and heterocyclic groups.
  • Alkyl refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 20 carbon atoms (“C 1-20 alkyl”). In some embodiments, an alkyl group has 1 to 10 carbon atoms (“C 1-10 alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C 1-9 alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C 1-8 alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C 1-7 alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C 1-6 alkyl”).
  • an alkyl group has 1 to 5 carbon atoms (“C 1-5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C 1-4 alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C 1-3 alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C 1-2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C 1 alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C 2-6 alkyl”).
  • C 1-6 alkyl groups include methyl (C 1 ), ethyl (C 2 ), n-propyl (C 3 ), isopropyl (C 3 ), n-butyl (C 4 ), tert-butyl (C 4 ), sec-butyl (C 4 ), iso-butyl (C 4 ), n-pentyl (C 5 ), 3-pentanyl (C 5 ), amyl (C 5 ), neopentyl (C 5 ), 3-methyl-2-butanyl (C 5 ), tertiary amyl (C 5 ), and n-hexyl (C 6 ).
  • alkyl groups include n-heptyl (C 7 ), n-octyl (C 8 ) and the like. Unless otherwise specified, each instance of an alkyl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted alkyl”) or substituted (a “substituted alkyl”) with one or more substituents.
  • the alkyl group is unsubstituted C 1-10 alkyl (such as unsubstituted C 1-6 alkyl, e.g., —CH 3 (Me), unsubstituted ethyl (Et), unsubstituted propyl (Pr, e.g., unsubstituted n-propyl (n-Pr), unsubstituted isopropyl (i-Pr)), unsubstituted butyl (Bu, e.g., unsubstituted n-butyl (n-Bu), unsubstituted tert-butyl (tert-Bu or t-Bu), unsubstituted sec-butyl (sec-Bu), unsubstituted isobutyl (i-Bu)).
  • the alkyl group is substituted C 1-10 alkyl (such as substituted C 1-6 alkyl, e.g., —CH 3 (Me), unsub
  • Alkenyl refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 20 carbon atoms, one or more carbon-carbon double bonds, and no triple bonds (“C 2-20 alkenyl”). In some embodiments, an alkenyl group has 2 to 10 carbon atoms (“C 2-10 alkenyl”). In some embodiments, an alkenyl group has 2 to 9 carbon atoms (“C 2-9 alkenyl”). In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C 2-8 alkenyl”). In some embodiments, an alkenyl group has 2 to 7 carbon atoms (“C 2-7 alkenyl”).
  • an alkenyl group has 2 to 6 carbon atoms (“C 2-6 alkenyl”). In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C 2-5 alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C 2-4 alkenyl”). In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C 2-3 alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“C 2 alkenyl”). The one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl).
  • Examples of C 2-4 alkenyl groups include ethenyl (C 2 ), 1-propenyl (C 3 ), 2-propenyl (C 3 ), 1-butenyl (C 4 ), 2-butenyl (C 4 ), butadienyl (C 4 ), and the like.
  • Examples of C 2-6 alkenyl groups include the aforementioned C 2-4 alkenyl groups as well as pentenyl (C 5 ), pentadienyl (C 5 ), hexenyl (C 6 ), and the like. Additional examples of alkenyl include heptenyl (C 7 ), octenyl (C 8 ), octatrienyl (C 8 ), and the like.
  • each instance of an alkenyl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) with one or more substituents.
  • the alkenyl group is unsubstituted C 2-10 alkenyl.
  • the alkenyl group is substituted C 2-10 alkenyl.
  • a C ⁇ C double bond for which the stereochemistry is not specified e.g., —CH ⁇ CHCH 3 or
  • Alkynyl refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 20 carbon atoms, one or more carbon-carbon triple bonds, and optionally one or more double bonds (“C 2-20 alkynyl”).
  • an alkynyl group has 2 to 10 carbon atoms (“C 2-10 alkynyl”).
  • an alkynyl group has 2 to 9 carbon atoms (“C 2-9 alkynyl”).
  • an alkynyl group has 2 to 8 carbon atoms (“C 2-8 alkynyl”).
  • an alkynyl group has 2 to 7 carbon atoms (“C 2-7 alkynyl”).
  • an alkynyl group has 2 to 6 carbon atoms (“C 2-6 alkynyl”). In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C 2-5 alkynyl”). In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C 2-4 alkynyl”). In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C 2-3 alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C 2 alkynyl”). The one or more carbon-carbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl).
  • Examples of C 2-4 alkynyl groups include, without limitation, ethynyl (C 2 ), 1-propynyl (C 3 ), 2-propynyl (C 3 ), 1-butynyl (C 4 ), 2-butynyl (C 4 ), and the like.
  • Examples of C 2-6 alkenyl groups include the aforementioned C 2-4 alkynyl groups as well as pentynyl (C 5 ), hexynyl (C 6 ), and the like. Additional examples of alkynyl include heptynyl (C 7 ), octynyl (C 8 ), and the like.
  • each instance of an alkynyl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted alkynyl”) or substituted (a “substituted alkynyl”) with one or more substituents.
  • the alkynyl group is unsubstituted C 2-10 alkynyl.
  • the alkynyl group is substituted C 2-10 alkynyl.
  • carbocyclyl refers to a radical of a non-aromatic cyclic hydrocarbon group having from 3 to 14 ring carbon atoms (“C 3-14 carbocyclyl”) and zero heteroatoms in the non-aromatic ring system.
  • a carbocyclyl group has 3 to 10 ring carbon atoms (“C 3-10 carbocyclyl”).
  • a carbocyclyl group has 3 to 8 ring carbon atoms (“C 3-8 carbocyclyl”).
  • a carbocyclyl group has 3 to 7 ring carbon atoms (“C 3-7 carbocyclyl”).
  • a carbocyclyl group has 3 to 6 ring carbon atoms (“C 3-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 4 to 6 ring carbon atoms (“C 4-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 6 ring carbon atoms (“C 5-6 carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C 5-10 carbocyclyl”).
  • Exemplary C 3-6 carbocyclyl groups include, without limitation, cyclopropyl (C 3 ), cyclopropenyl (C 3 ), cyclobutyl (C 4 ), cyclobutenyl (C 4 ), cyclopentyl (C 5 ), cyclopentenyl (C 5 ), cyclohexyl (C 6 ), cyclohexenyl (C 6 ), cyclohexadienyl (C 6 ), and the like.
  • Exemplary C 3-8 carbocyclyl groups include, without limitation, the aforementioned C 3-6 carbocyclyl groups as well as cycloheptyl (C 7 ), cycloheptenyl (C 7 ), cycloheptadienyl (C 7 ), cycloheptatrienyl (C 7 ), cyclooctyl (C 8 ), cyclooctenyl (C 8 ), bicyclo[2.2.1]heptanyl (C 7 ), bicyclo[2.2.2]octanyl (C 8 ), and the like.
  • Exemplary C 3-10 carbocyclyl groups include, without limitation, the aforementioned C 3-8 carbocyclyl groups as well as cyclononyl (C 9 ), cyclononenyl (C 9 ), cyclodecyl (C 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C 9 ), decahydronaphthalenyl (C 10 ), spiro[4.5]decanyl (C 10 ), and the like.
  • the carbocyclyl group is either monocyclic (“monocyclic carbocyclyl”) or polycyclic (e.g., containing a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic carbocyclyl”) or tricyclic system (“tricyclic carbocyclyl”)) and can be saturated or can contain one or more carbon-carbon double or triple bonds.
  • Carbocyclyl also includes ring systems wherein the carbocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups wherein the point of attachment is on the carbocyclyl ring, and in such instances, the number of carbons continue to designate the number of carbons in the carbocyclic ring system.
  • each instance of a carbocyclyl group is independently unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a “substituted carbocyclyl”) with one or more substituents.
  • the carbocyclyl group is an unsubstituted C 3-14 carbocyclyl.
  • the carbocyclyl group is a substituted C 3-14 carbocyclyl.
  • “carbocyclyl” is a monocyclic, saturated carbocyclyl group having from 3 to 14 ring carbon atoms (“C 3-14 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 10 ring carbon atoms (“C 3-10 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C 3-8 cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C 3-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 4 to 6 ring carbon atoms (“C 4-6 cycloalkyl”).
  • a cycloalkyl group has 5 to 6 ring carbon atoms (“C 5-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C 5-10 cycloalkyl”). Examples of C 5-6 cycloalkyl groups include cyclopentyl (C 5 ) and cyclohexyl (C 5 ). Examples of C 3-6 cycloalkyl groups include the aforementioned C 5-6 cycloalkyl groups as well as cyclopropyl (C 3 ) and cyclobutyl (C 4 ).
  • C 3-8 cycloalkyl groups include the aforementioned C 3-6 cycloalkyl groups as well as cycloheptyl (C 7 ) and cyclooctyl (C 8 ).
  • each instance of a cycloalkyl group is independently unsubstituted (an “unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”) with one or more substituents.
  • the cycloalkyl group is an unsubstituted C 3-14 cycloalkyl.
  • the cycloalkyl group is a substituted C 3-14 cycloalkyl.
  • Heterocyclyl or “heterocyclic” refers to a radical of a 3- to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“3-10 membered heterocyclyl”).
  • the point of attachment can be a carbon or nitrogen atom, as valency permits.
  • a heterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”) or a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”), and can be saturated or can be partially unsaturated.
  • Heterocyclyl bicyclic ring systems can include one or more heteroatoms in one or both rings.
  • Heterocyclyl also includes ring systems wherein the heterocyclic ring, as defined above, is fused with one or more carbocyclyl groups wherein the point of attachment is either on the carbocyclyl or heterocyclic ring, or ring systems wherein the heterocyclic ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclic ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclic ring system.
  • each instance of heterocyclyl is independently optionally substituted, i.e., unsubstituted (an “unsubstituted heterocyclyl”) or substituted (a “substituted heterocyclyl”) with one or more substituents.
  • the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. In certain embodiments, the heterocyclyl group is substituted 3-10 membered heterocyclyl.
  • a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon (“5-10 membered heterocyclyl”).
  • a heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heterocyclyl”).
  • a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”).
  • the 5-6 membered heterocyclyl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the 5-6 membered heterocyclyl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the 5-6 membered heterocyclyl has one ring heteroatom selected from nitrogen, oxygen, and sulfur.
  • Exemplary 3-membered heterocyclyl groups containing one heteroatom include, without limitation, azirdinyl, oxiranyl, and thiiranyl.
  • Exemplary 4-membered heterocyclyl groups containing one heteroatom include, without limitation, azetidinyl, oxetanyl and thietanyl.
  • Exemplary 5-membered heterocyclyl groups containing one heteroatom include, without limitation, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione.
  • Exemplary 5-membered heterocyclyl groups containing two heteroatoms include, without limitation, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one.
  • Exemplary 5-membered heterocyclyl groups containing three heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl.
  • Exemplary 6-membered heterocyclyl groups containing one heteroatom include, without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl.
  • Exemplary 6-membered heterocyclyl groups containing two heteroatoms include, without limitation, piperazinyl, morpholinyl, dithianyl, and dioxanyl.
  • Exemplary 6-membered heterocyclyl groups containing three heteroatoms include, without limitation, triazinyl.
  • Exemplary 7-membered heterocyclyl groups containing one heteroatom include, without limitation, azepanyl, oxepanyl and thiepanyl.
  • Exemplary 8-membered heterocyclyl groups containing one heteroatom include, without limitation, azocanyl, oxecanyl and thiocanyl.
  • Exemplary 5-membered heterocyclyl groups fused to a C 6 aryl ring include, without limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and the like.
  • Exemplary 6-membered heterocyclyl groups fused to an aryl ring include, without limitation, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.
  • Aryl refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 p electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (“C 6-14 aryl”).
  • an aryl group has six ring carbon atoms (“C 6 aryl”; e.g., phenyl).
  • an aryl group has ten ring carbon atoms (“C 10 aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl).
  • an aryl group has fourteen ring carbon atoms (“C 14 aryl”; e.g., anthracyl).
  • Aryl also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system.
  • each instance of an aryl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents.
  • the aryl group is unsubstituted C 6-14 aryl.
  • the aryl group is substituted C 6-14 aryl.
  • Alkyl is a subset of alkyl and aryl and refers to an optionally substituted alkyl group substituted by an optionally substituted aryl group. In certain embodiments, the aralkyl is optionally substituted benzyl. In certain embodiments, the aralkyl is benzyl. In certain embodiments, the aralkyl is optionally substituted phenethyl. In certain embodiments, the aralkyl is phenethyl.
  • Heteroaryl refers to a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 p electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur (“5-10 membered heteroaryl”).
  • heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits.
  • Heteroaryl bicyclic ring systems can include one or more heteroatoms in one or both rings.
  • Heteroaryl includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heteroaryl ring system. “Heteroaryl” also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused (aryl/heteroaryl) ring system.
  • Bicyclic heteroaryl groups wherein one ring does not contain a heteroatom e.g., indolyl, quinolinyl, carbazolyl, and the like
  • the point of attachment can be on either ring, i.e., either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl).
  • a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”).
  • a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”).
  • a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”).
  • the 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the 5-6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur.
  • each instance of a heteroaryl group is independently optionally substituted, i.e., unsubstituted (an “unsubstituted heteroaryl”) or substituted (a “substituted heteroaryl”) with one or more substituents.
  • the heteroaryl group is unsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is substituted 5-14 membered heteroaryl.
  • Exemplary 5-membered heteroaryl groups containing one heteroatom include, without limitation, pyrrolyl, furanyl, and thiophenyl.
  • Exemplary 5-membered heteroaryl groups containing two heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl.
  • Exemplary 5-membered heteroaryl groups containing three heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl.
  • Exemplary 5-membered heteroaryl groups containing four heteroatoms include, without limitation, tetrazolyl.
  • Exemplary 6-membered heteroaryl groups containing one heteroatom include, without limitation, pyridinyl.
  • Exemplary 6-membered heteroaryl groups containing two heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl.
  • Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively.
  • Exemplary 7-membered heteroaryl groups containing one heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl.
  • Exemplary 5,6-bicyclic heteroaryl groups include, without limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl.
  • Exemplary 6,6-bicyclic heteroaryl groups include, without limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.
  • Heteroaralkyl is a subset of alkyl and heteroaryl and refers to an optionally substituted alkyl group substituted by an optionally substituted heteroaryl group.
  • Partially unsaturated refers to a group that includes at least one double or triple bond.
  • a “partially unsaturated” ring system is further intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic groups (e.g., aryl or heteroaryl groups) as herein defined.
  • aromatic groups e.g., aryl or heteroaryl groups
  • saturated refers to a group that does not contain a double or triple bond, i.e., contains all single bonds.
  • Alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups, which are divalent bridging groups are further referred to using the suffix -ene, e.g., alkylene, alkenylene, alkynylene, carbocyclylene, heterocyclylene, arylene, and heteroarylene.
  • Alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups are optionally substituted (e.g., “substituted” or “unsubstituted” alkyl, “substituted” or “unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl, “substituted” or “unsubstituted” carbocyclyl, “substituted” or “unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or “substituted” or “unsubstituted” heteroaryl group) if not otherwise provided explicitly.
  • substituted means that at least one hydrogen present on a group (e.g., a carbon or nitrogen atom) is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction.
  • a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position.
  • substituted is contemplated to include substitution with all permissible substituents of organic compounds, any of the substituents described herein that results in the formation of a stable compound.
  • the present invention contemplates any and all such combinations in order to arrive at a stable compound.
  • heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.
  • Exemplary carbon atom substituents include, but are not limited to, halogen, —CN, —NO 2 , —N 3 , —SO 2 H, —SO 3 H, —OH, —OR aa , —ON(R bb ) 2 , —N(R bb ) 2 , —N(R bb ) 3 + X ⁇ , —N(OR cc )R bb , —SH, —SR aa , —SSR cc , —C( ⁇ O)R aa , —CO 2 H, —CHO, —C(OR cc ) 2 , —CO 2 R aa , —OC( ⁇ O)R aa , —OCO 2 R aa , —C( ⁇ O)N(R bb ) 2 , —OC( ⁇ O)N(R bb ) 2 , —NR bb C
  • each instance of R aa is, independently, selected from C 1-10 alkyl, C 1-10 perhaloalkyl, C 2-10 alkenyl, C 2-10 alkynyl, heteroC 1-10 alkyl, heteroC 2-10 alkenyl, heteroC 2-10 alkynyl, C 3-10 carbocyclyl, 3-14 membered heterocyclyl, C 6-14 aryl, and 5-14 membered heteroaryl, or two R aa groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R dd groups;
  • each instance of R bb is, independently, selected from hydrogen, —OH, —OR aa , —N(R cc ) 2 , —CN, —C( ⁇ O)R aa , —C( ⁇ O)N(R cc ) 2 , —CO 2 R aa , —SO 2 R aa , —C( ⁇ NR cc )OR aa , —C( ⁇ NR cc )N(R cc ) 2 , —SO 2 N(R cc ) 2 , —SO 2 R cc , —SO 2 OR cc , —SOR aa , —C( ⁇ S)N(R cc ) 2 , —C( ⁇ O)SR cc , —C( ⁇ S)SR cc , —P( ⁇ O)(R aa ) 2 , —P( ⁇ O)(OR cc ) 2
  • each instance of R cc is, independently, selected from hydrogen, C 1-10 alkyl, C 1-10 perhaloalkyl, C 2-10 alkenyl, C 2-10 alkynyl, heteroC 1-10 alkyl, heteroC 2-10 alkenyl, heteroC 2-10 alkynyl, C 3-10 carbocyclyl, 3-14 membered heterocyclyl, C 6-14 aryl, and 5-14 membered heteroaryl, or two R cc groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R dd groups;
  • each instance of R dd is, independently, selected from halogen, —CN, —NO 2 , —N 3 , —SO 2 H, —SO 3 H, —OH, —OR ee , —ON(R ff ) 2 , —N(R ff ) 2 , —N(R ff ) 3 + X ⁇ , —N(OR ee )R ff , —SH, —SR ee , —SSR ee , —C( ⁇ O)R ee , —CO 2 H, —CO 2 R ee , —OC( ⁇ O)R ee , —OCO 2 R ee , —C( ⁇ O)N(R ff ) 2 , —OC( ⁇ O)N(R ff ) 2 , —NR ff C( ⁇ O)R ee , —NR ff CO 2 R
  • each instance of R ee is, independently, selected from C 1-6 alkyl, C 1-6 perhaloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, heteroC 1-6 alkyl, heteroC 2-6 alkenyl, heteroC 2-6 alkynyl, C 3-10 carbocyclyl, C 6-10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R gg groups;
  • each instance of R ff is, independently, selected from hydrogen, C 1-6 alkyl, C 1-6 perhaloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, heteroC 1-6 alkyl, heteroC 2-6 alkenyl, heteroC 2-6 alkynyl, C 3-10 carbocyclyl, 3-10 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl, or two R ff groups are joined to form a 3-10 membered heterocyclyl or 5-10 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R gg groups; and
  • each instance of R gg is, independently, halogen, —CN, —NO 2 , —N 3 , —SO 2 H, —SO 3 H, —OH, —OC 1-6 alkyl, —ON(C 1-6 alkyl) 2 , —N(C 1-6 alkyl) 2 , —N(C 1-6 alkyl) 3 + X ⁇ , —NH(C 1-6 alkyl) 2 + X ⁇ , —NH 2 (C 1-6 alkyl) + X ⁇ , —NH 3 + X ⁇ , —N(OC 1-6 alkyl)(C 1-6 alkyl), —N(OH)(C 1-6 alkyl), —NH(OH), —SH, —SC 1-6 alkyl, —SS(C 1-6 alkyl), —C( ⁇ O)(C 1-6 alkyl), —CO 2 H, —CO 2 (C 1-6 alkyl), —OC( ⁇ O)
  • the carbon atom substituents are independently halogen, substituted or unsubstituted C 1-6 alkyl, or —OR aa .
  • a “counterion” or “anionic counterion” is a negatively charged group associated with a positively charged group in order to maintain electronic neutrality.
  • An anionic counterion may be monovalent (i.e., including one formal negative charge).
  • An anionic counterion may also be multivalent (i.e., including more than one formal negative charge), such as divalent or trivalent.
  • Exemplary counterions include halide ions (e.g., F ⁇ , Cl ⁇ , Br ⁇ , I ⁇ ), NO 3 ⁇ , ClO 4 ⁇ , OH ⁇ , H 2 PO 4 ⁇ , HCO 3 ⁇ , HSO 4 ⁇ , sulfonate ions (e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonic acid-2-sulfonate, and the like), carboxylate ions (e.g., acetate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, gluconate, and the like), BF 4
  • Exemplary counterions which may be multivalent include CO 3 2 ⁇ , HPO 4 2 ⁇ , PO 4 3 ⁇ , B 4 O 7 2 ⁇ , SO 4 2 ⁇ , S 2 O 3 2 ⁇ , carboxylate anions (e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelate, sebacate, salicylate, phthalates, aspartate, glutamate, and the like), and carboranes.
  • carboxylate anions e.g., tartrate, citrate, fumarate, maleate, malate, malonate, gluconate, succinate, glutarate, adipate, pimelate, suberate, azelate, sebacate, salicylate, phthalates, aspartate, glutamate, and the like
  • carboranes e.g., tartrate, citrate, fumarate, maleate, mal
  • Halo or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro, —Cl), bromine (bromo, —Br), or iodine (iodo, —I).
  • hydroxyl refers to the group —OH.
  • substituted hydroxyl or “substituted hydroxyl,” by extension, refers to a hydroxyl group wherein the oxygen atom directly attached to the parent molecule is substituted with a group other than hydrogen, and includes groups selected from —OR aa , —ON(R bb ) 2 , —OC( ⁇ O)SR aa , —OC( ⁇ O)R aa , —OCO 2 R aa , —OC( ⁇ O)N(R bb ) 2 , —OC( ⁇ NR bb )R aa , —OC( ⁇ NR bb )OR aa , —OC( ⁇ NR bb )N(R bb ) 2 , —OS( ⁇ O)R aa , —OSO 2 R aa , —OSi(R aa ) 3 , —
  • amino refers to the group —NH 2 .
  • substituted amino by extension, refers to a monosubstituted amino, a disubstituted amino, or a trisubstituted amino. In certain embodiments, the “substituted amino” is a monosubstituted amino or a disubstituted amino group.
  • “Acyl” refers to a moiety selected from the group consisting of —C( ⁇ O)R aa , —CHO, —CO 2 R aa , —C( ⁇ O)N(R bb ) 2 , —C( ⁇ NR bb )R aa , —C( ⁇ NR bb )OR aa , —C( ⁇ NR bb )N(R bb ) 2 , —C( ⁇ O)NR bb SO 2 R aa , —C( ⁇ S)N(R bb ) 2 , —C( ⁇ O)SR aa , or —C( ⁇ S)SR aa , wherein R aa and R bb are as defined herein.
  • Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quaternary nitrogen atoms.
  • Exemplary nitrogen atom substituents include, but are not limited to, hydrogen, —OH, —OR aa , —N(R cc ) 2 , —CN, —C( ⁇ O)R aa , —C( ⁇ O)N(R cc ) 2 , —CO 2 R aa , —SO 2 R aa , —C( ⁇ NR bb )R aa , —C( ⁇ NR cc )OR aa , —C( ⁇ NR cc )N(R cc ) 2 , —SO 2 N(R cc ) 2 , —SO 2 R cc , —SO 2 OR cc , —SOR aa , —C( ⁇ S)N(R
  • the substituent present on a nitrogen atom is a nitrogen protecting group (also referred to as an amino protecting group).
  • Nitrogen protecting groups include, but are not limited to, —OH, —OR aa , —N(R cc ) 2 , —C( ⁇ O)R aa , C( ⁇ O)N(R cc ) 2 , —CO 2 R aa , —SO 2 R aa , —C( ⁇ NR cc )R aa , —C( ⁇ NR cc )OR aa , —C( ⁇ NR cc )N(R cc ) 2 , —SO 2 N(R cc ) 2 , —SO 2 R cc , —SO 2 OR cc , —SOR aa , —C( ⁇ S)N(R cc ) 2 , —C( ⁇ O)SR cc , —C( ⁇ S)
  • Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis , T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, incorporated herein by reference.
  • nitrogen protecting groups such as amide groups (e.g., —C( ⁇ O)R aa ) include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N′-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitro
  • Nitrogen protecting groups such as carbamate groups include, but are not limited to, methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate
  • Nitrogen protecting groups such as sulfonamide groups include, but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide
  • Ts p-toluenesulfonamide
  • nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivative, N′-p-toluenesulfonylaminoacyl derivative, N-phenylaminothioacyl derivative, N-benzoylphenylalanyl derivative, N-acetylmethionine derivative, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-
  • the substituent present on an oxygen atom is an oxygen protecting group (also referred to herein as an “hydroxyl protecting group”).
  • Oxygen protecting groups include, but are not limited to, —R aa , —N(R bb ) 2 , —C( ⁇ O)SR aa , —C( ⁇ O)R aa , —CO 2 R aa , —C( ⁇ O)N(R bb ) 2 , —C( ⁇ NR bb )R aa , —C( ⁇ NR bb )OR aa , —C( ⁇ NR bb )N(R bb ) 2 , —S( ⁇ O)R aa , —SO 2 R aa , —Si(R aa ) 3 , —P(R cc ) 2 , —P(R cc ) 3 + X ⁇ , —P(OR cc
  • Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, incorporated herein by reference.
  • oxygen protecting groups include, but are not limited to, methyl, methoxylmethyl (MOM), tert-butoxycarbonyl, methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethyl silyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethyl silyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydr
  • the substituent present on a sulfur atom is a sulfur protecting group (also referred to as a “thiol protecting group”).
  • Sulfur protecting groups include, but are not limited to, —R aa , —N(R bb ) 2 , —C( ⁇ O)SR aa , —C( ⁇ O)R aa , —CO 2 R aa , —C( ⁇ O)N(R bb ) 2 , —C( ⁇ NR bb )R aa , —C( ⁇ NR bb )OR aa , —C( ⁇ NR bb )N(R bb ) 2 , —S( ⁇ O)R aa , —SO 2 R aa , —Si(R aa ) 3 , —P(R cc ) 2 , —P(R cc ) 3 + X ⁇ , —P(OR c
  • Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3 rd edition, John Wiley & Sons, 1999, incorporated herein by reference.
  • pharmaceutically acceptable salt refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference.
  • Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases.
  • Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid
  • organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
  • Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N + (C 1-4 alkyl) 4 ⁇ salts.
  • Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.
  • solvate refers to forms of the compound, or a salt thereof, that are associated with a solvent, usually by a solvolysis reaction. This physical association may include hydrogen bonding.
  • Conventional solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, and the like.
  • the compounds of Formula (I′) e.g., Formula (I)), (II), or (III) may be prepared, e.g., in crystalline form, and may be solvated.
  • Suitable solvates include pharmaceutically acceptable solvates and further include both stoichiometric solvates and non-stoichiometric solvates.
  • the solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated in the crystal lattice of a crystalline solid.
  • “Solvate” encompasses both solution-phase and isolable solvates.
  • Representative solvates include hydrates, ethanolates, and methanolates.
  • hydrate refers to a compound that is associated with water.
  • the number of the water molecules contained in a hydrate of a compound is in a definite ratio to the number of the compound molecules in the hydrate. Therefore, a hydrate of a compound may be represented, for example, by the general formula R.x H 2 O, wherein R is the compound and wherein x is a number greater than 0.
  • a given compound may form more than one type of hydrate, including, e.g., monohydrates (x is 1), lower hydrates (x is a number greater than 0 and smaller than 1, e.g., hemihydrates (R.0.5H 2 O)), and polyhydrates (x is a number greater than 1, e.g., dihydrates (R.2H 2 O) and hexahydrates (R.6H 2 O)).
  • monohydrates x is 1
  • lower hydrates x is a number greater than 0 and smaller than 1, e.g., hemihydrates (R.0.5H 2 O)
  • polyhydrates x is a number greater than 1, e.g., dihydrates (R.2H 2 O) and hexahydrates (R.6H 2 O)
  • tautomers or “tautomeric” refers to two or more interconvertible compounds resulting from at least one formal migration of a hydrogen atom and at least one change in valency (e.g., a single bond to a double bond, a triple bond to a single bond, or vice versa).
  • the exact ratio of the tautomers depends on several factors, including temperature, solvent, and pH. Tautomerizations (i.e., the reaction providing a tautomeric pair) may catalyzed by acid or base.
  • Exemplary tautomerizations include keto-to-enol, amide-to-imide, lactam-to-lactim, enamine-to-imine, and enamine-to-(a different enamine) tautomerizations.
  • Tautomeric forms may be relevant to the attainment of the optimal chemical reactivity and biological activity of a compound of interest.
  • stereoisomers that are not mirror images of one another are termed “diastereomers” and those that are non-superimposable mirror images of each other are termed “enantiomers”.
  • enantiomers When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible.
  • An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or ( ⁇ )-isomers respectively).
  • a chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a “racemic mixture”.
  • Polymorph refers to a particular polymorphic variant of a given compound. Polymorphism is the ability of a solid substance of a given chemical composition to exist in more than one form or crystalline structure. Polymorphism can exist as a result of differences in crystal packing (packing polymorphism), conformational differences (conformational polymorphism), or changes due to co-crystalization with other chemical entities (pseudopolymorphism). Polymorphism is an important aspect of pharmaceutical development, in which case drugs typically receive regulatory approval for only a single form. Distinct polymorphic forms frequently vary considerably in terms of their physical properties. Altered dissolution rates, thermal stability, and hygroscopicity are frequently observed.
  • prodrugs refers to compounds that have cleavable groups and become by solvolysis or under physiological conditions the compounds of Formula (I′) (e.g., Formula (I)), (II), or (III), which are pharmaceutically active in vivo.
  • Such examples include, but are not limited to, ester derivatives and the like.
  • Other derivatives of the compounds of this invention have activity in both their acid and acid derivative forms, but in the acid sensitive form often offers advantages of solubility, tissue compatibility, or delayed release in the mammalian organism (see, Bundgard, H., Design of Prodrugs , pp. 7-9, 21-24, Elsevier, Amsterdam 1985).
  • Prodrugs include acid derivatives well known to practitioners of the art, such as, for example, esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a substituted or unsubstituted amine, or acid anhydrides, or mixed anhydrides.
  • Simple aliphatic or aromatic esters, amides, and anhydrides derived from acidic groups pendant on the compounds of this invention are particular prodrugs.
  • double ester type prodrugs such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkylesters.
  • C 1 to C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, aryl, C 7 -C 12 substituted aryl, and C 7 -C 12 arylalkyl esters of the compounds of Formula (I′) e.g., Formula (I)), (II), or (III) may be preferred.
  • a “subject” to which administration is contemplated refers to a human (i.e., male or female of any age group, e.g., pediatric subject (e.g., infant, child, or adolescent) or adult subject (e.g., young adult, middle-aged adult, or senior adult)) or non-human animal.
  • the non-human animal is a mammal (e.g., primate (e.g., cynomolgus monkey or rhesus monkey), commercially relevant mammal (e.g., cattle, pig, horse, sheep, goat, cat, or dog), or bird (e.g., commercially relevant bird, such as chicken, duck, goose, or turkey)).
  • primate e.g., cynomolgus monkey or rhesus monkey
  • commercially relevant mammal e.g., cattle, pig, horse, sheep, goat, cat, or dog
  • bird e.g., commercially relevant bird, such as
  • the non-human animal is a fish, reptile, or amphibian.
  • the non-human animal may be a male or female at any stage of development.
  • the non-human animal may be a transgenic animal or genetically engineered animal.
  • a “patient” refers to a human subject in need of treatment of a disease.
  • the subject may also be a plant.
  • the plant is a land plant.
  • the plant is a non-vascular land plant.
  • the plant is a vascular land plant.
  • the plant is a seed plant.
  • the plant is a cultivated plant.
  • the plant is a dicot.
  • the plant is a monocot.
  • the plant is a flowering plant.
  • the plant is a cereal plant, e.g., maize, corn, wheat, rice, oat, barley, rye, or millet.
  • the plant is a legume, e.g., a bean plant, e.g., soybean plant.
  • the plant is a tree or shrub.
  • administer refers to implanting, absorbing, ingesting, injecting, inhaling, or otherwise introducing an inventive compound, or a pharmaceutical composition thereof.
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a microbial infection (e.g., a bacterial infection or mycobacterial infection).
  • treatment may be administered after one or more signs or symptoms have developed or have been observed.
  • treatment may be administered in the absence of signs or symptoms of the disease or condition.
  • treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of exposure to microorganisms, in light of a history of symptoms, and/or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example, to delay and/or prevent recurrence.
  • an “effective amount” of a compound described herein refers to an amount sufficient to elicit the desired biological response.
  • An effective amount of a compound described herein may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health of the subject.
  • an effective amount is a therapeutically effective amount.
  • an effective amount is a prophylactic treatment.
  • an effective amount is the amount of a compound described herein in a single dose.
  • an effective amount is the combined amounts of a compound described herein in multiple doses.
  • an effective amount is an amount effective for inhibiting the growth of a microorganism, for inhibiting the reproduction of a microorganism, or for killing a microorganism. In certain embodiments, an effective amount is an amount effective for inhibiting the formation of a biofilm, for inhibiting the growth of a biofilm, for reducing a biofilm, or for clearing a biofilm. In certain embodiments, an effective amount is an amount effective for disinfecting a surface (e.g., killing at least 80%, at least 90%, at least 99%, at least 99.9%, or at least 99.99% of the microorganisms on the surface). In certain embodiments, an effective amount is an amount effective for killing a persister cell.
  • a “therapeutically effective amount” of a compound of Formula (I′) is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or to delay or minimize one or more symptoms associated with the condition.
  • a therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the condition.
  • the term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of the condition, or enhances the therapeutic efficacy of another therapeutic agent.
  • a therapeutically effective amount is effective for treating a microbial infection (e.g., a bacterial infection or mycobacterial infection) in a subject, for inhibiting the growth and/or reproduction of a microorganism (e.g., a bacterium), for killing a microorganism (e.g., a bacterium), for inhibiting the formation and/or growth of a biofilm, for reducing or clearing a biofilm, and/or for disinfecting a surface.
  • a microbial infection e.g., a bacterial infection or mycobacterial infection
  • a “prophylactically effective amount” of a compound of Formula (I′) is an amount sufficient to prevent a condition, or one or more symptoms associated with the condition or prevent its recurrence.
  • a prophylactically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other agents, which provides a prophylactic benefit in the prevention of the condition.
  • the term “prophylactically effective amount” can encompass an amount that improves overall prophylaxis or enhances the prophylactic efficacy of another prophylactic agent.
  • a prophylactically effective amount is effective for preventing a microbial infection (e.g., a bacterial infection or mycobacterial infection) in a subject, for inhibiting the growth and/or reproduction of a microorganism (e.g., a bacterium), for killing a microorganism (e.g., a bacterium), for inhibiting the formation and/or growth of a biofilm, for reducing or clearing a biofilm, and/or for disinfecting a surface.
  • a microbial infection e.g., a bacterial infection or mycobacterial infection
  • inhibitor refers to the ability of a compound to reduce, slow, halt, or prevent activity of a particular biological process (e.g., the growth or reproduction) of a microorganism (e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite) relative to vehicle.
  • a microorganism e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite
  • MIC minimum inhibitory concentration
  • a microorganism e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite
  • overnight e.g., about 16 to about 20 hours, or about 16 to about 18 hours
  • half maximal inhibitory concentration or “IC 50 ” of a compound refers to the concentration of the compound that inhibits the growth of half of an inoculum of a microorganism (e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite).
  • a microorganism e.g., a bacterium, mycobacterium , archaeon, protist, fungus, or parasite.
  • microorganism refers to a microscopic organism, which may be a single-cell or multicellular organism.
  • the microorganism is a bacterium, mycobacterium , archaeon, protist (e.g., protozoon, alga), fungus (e.g., yeast, mold), or parasite.
  • the microorganism is a bacterium.
  • the length or diameter of a microorganism is at most about 10 cm, at most about 1 cm, at most about 1 mm, at most about 100 ⁇ m, at most about 10 ⁇ m, at most about 1 ⁇ m, at most about 100 nm, or at most about 10 nm. In certain embodiments, the length or diameter of a microorganism is at most about 10 ⁇ m.
  • biofilm refers to a group of microorganisms (e.g., bacteria) in which cells of the microorganisms stick to each other on a surface. These adherent cells are frequently embedded within a self-produced matrix of extracellular polymeric substance (EPS).
  • EPS extracellular polymeric substance
  • Biofilms may form on living or non-living surfaces and can be prevalent in natural, industrial, and hospital settings. The cells growing in a biofilm are physiologically distinct from planktonic cells of the same microorganism, which are single-cells that may float or swim in a liquid medium. Biofilms have been found to be involved in a wide variety of microbial infections. Biofilms are formed by numerous Gram-negative and Gram-positive bacterial species. Non-limiting examples include Bacillus spp, Staphylococcus spp, Pseudomonas spp, and Acinetobacter spp.
  • microbial warfare refers to a first microorganism producing a substance (e.g., an antibiotic) that is toxic to a second microorganism but is not toxic or less toxic, compared to the second microorganism, to the first microorganism.
  • a second microorganism in close proximity to the first microorganism contacts the substance the growth and/or reproduction of the second microorganism may be inhibited, or the second microorganism may be killed.
  • the first microorganism may gain a competitive advantage over the second microorganism in close proximity to the first microorganism in terms of survival, growth, and/or reproduction.
  • tissue sample refers to any sample including tissue samples (such as tissue sections and needle biopsies of a tissue); cell samples (e.g., cytological smears (such as Pap or blood smears) or samples of cells obtained by microdissection); samples of whole organisms (such as samples of yeasts or bacteria); or cell fractions, fragments or organelles (such as obtained by lysing cells and separating the components thereof by centrifugation or otherwise).
  • tissue samples such as tissue sections and needle biopsies of a tissue
  • cell samples e.g., cytological smears (such as Pap or blood smears) or samples of cells obtained by microdissection) or samples of cells obtained by microdissection
  • samples of whole organisms such as samples of yeasts or bacteria
  • cell fractions, fragments or organelles such as obtained by lysing cells and separating the components thereof by centrifugation or otherwise.
  • biological samples include blood, serum, urine, semen, fecal matter, cerebrospinal fluid, interstitial fluid, mucus, tears, sweat, pus, biopsied tissue (e.g., obtained by a surgical biopsy or needle biopsy), nipple aspirates, milk, vaginal fluid, saliva, swabs (such as buccal swabs), or any material containing biomolecules that is derived from a first biological sample.
  • Biological samples also include those biological samples that are transgenic, such as transgenic oocyte, sperm cell, blastocyst, embryo, fetus, donor cell, or cell nucleus.
  • planktonic refers to any of the group of passively floating, drifting, or somewhat motile organisms occurring in a liquid medium (e.g., an aqueous solution). This group includes, but is not limited to, microscopic bacteria, algae, or protozoa.
  • FIG. 1 shows marine phenazine antibiotic 201 and an MRSA biofilm-eradicating agent, halogenated phenazine 202, as unique starting points to target persistent bacteria.
  • FIG. 2A shows exemplary results of Calgary Biofilm Device assays of 214 and vancomycin (“Vanco.”) against MRSA-2.
  • FIG. 2B shows Live/Dead staining of MRSA-2 biofilms following treatment with 214.
  • FIG. 2C shows Life/Dead staining of MRSA-2 biofilms following treatment with 213.
  • FIG. 3 shows eradication of MRSA-2 persister cells in a non-biofilm (stationary) culture.
  • Compound 214 demonstrated effective persister cell killing compared to lead MRSA antibiotics and QAC-10.
  • vancomycin denotes vancomycin.
  • Dapto denotes daptomycin.
  • FIG. 4 shows a flowchart illustrating a Calgary Biofilm Device (CBD) assay.
  • FIG. 5A shows MRSA-2 persister cell kill kinetics (killing of stationary cultures) of select compounds.
  • FIG. 5B shows additional MRSA-2 persister cell kill kinetics (killing of stationary cultures) of select compounds.
  • vanco denotes vancomycin.
  • FIG. 6A compound 202.
  • FIG. 6B compound 209.
  • FIG. 6C compound 210.
  • FIG. 6D compound 213.
  • FIG. 6E compound 214.
  • FIG. 7 shows synthetic analogues of marine phenazine antibiotic (1) that were evaluated during these investigations.
  • FIGS. 8A to 8D shows Schemes 3A to 3D: ( FIG. 8A (Scheme 3A)) mono- and di-halogenation routes to HP analogues 2-21, ( FIG. 8B (Scheme 3B)) Wohl-Aue synthesis of HP 18, ( FIG. 8C (Scheme 3C)) Suzuki Route to 4-butyl HP 22, and ( FIG. 8D (Scheme 3D))O-allylation/Clainsen Rearrangement leading to HPs 23-24.
  • FIG. 9 shows Calgary Biofilm Device assays to quantify planktonic (MBC) and biofilm (MBEC) killing efficiencies against MRSA-2, MRSE and VRE.
  • FIG. 10 shows biofilm cell killing (CFU/mL) for HP 14 obtained by colony counts from Calgary Biofilm Device pegs.
  • FIG. 11 shows live/dead staining of established MRSE 35984 biofilms treated with HP 14.
  • FIG. 12 shows structure-activity relationships (MIC/MBEC against MRSA-2) of all 29 HP analogues investigated during these studies. Diverse sub-classes of HP analogues with antibacterial and biofilm eradication activities against MRSA-2.
  • FIG. 13 shows detailed structure-activity relationships and antibacterial profiles of select halogenated phenazine analogues against MRSA, MRSE, VRE and MtB.
  • FIGS. 14A to 14D show UV-Vis analysis of metal chelation with 2 and 29.
  • FIG. 14A shows HP 2 binding copper(II) resulting in a loss of absorbance due to complex precipitation (insoluble).
  • FIG. 14B shows halogenated quinoline 29 binding copper(II) results in a shift in absorbance, which remains soluble.
  • FIG. 14D shows 29 binding iron(II) resulting in a shift in absorbance.
  • FIG. 15 shows HPs 2 and 29 complex formation with Cu(II) and Fe(II) determined by spectrophotometrically quantifying the concentration of free phenazine in solution following incubation with varying equivalents of CuSO 4
  • FIG. 16 shows HPs 2 and 29 complex formation with Cu(II) and Fe(II).
  • concentrations of free HP 2 in solution were determined from a calibration curve of serial dilutions of HP 2 in dimethyl sulfoxide.
  • FIG. 17 shows biofilm cell killing (CFU/mL) for HP 14 and 16 obtained by colony counts from Calgary Biofilm Device pegs.
  • FIGS. 18A to 18D show the kill kinetics of exponential growth cultures of S. aureus ( FIG. 18A ), MRSA-2 ( FIG. 18B ), S. epidermidis ( FIG. 18C ), and E. faecium ( FIG. 18D ) (rapidly-dividing bacteria) of select compounds.
  • FIGS. 20A to 20C show spectrophotometric determination of dissociation constants for HP 2.
  • FIG. 20A shows pH-dependent spectra scan results of HP 2.
  • FIG. 20B shows absorbance vs. pH curves of HP 2.
  • FIG. 20C shows pH vs. log [A ⁇ /HA] relationship of HP 2.
  • FIG. 21 shows biolfilm eradication against MRSA-2.
  • vancomycin denotes vancomycin
  • Cipro denotes ciprofloxacin
  • Dapto denotes daptomycin
  • Linezo denotes linezolid.
  • FIG. 22 shows biolfilm eradication against MRSA-2.
  • FIG. 23 shows biolfilm eradication against MRSA-2.
  • FIG. 24 shows biolfilm eradication against MRSA-2 (top panel) and biofilm eradication against MRSA BAA-44 (bottom panel).
  • FIG. 25 shows biolfilm eradication against MRSA BAA-1707.
  • FIG. 26 shows biolfilm eradication against S. epidermidis (MRSE 35984).
  • FIG. 27 shows biolfilm eradication against E. faecium (VRE 700221).
  • FIG. 28 shows a MIC assay against MRSA-2.
  • FIG. 29 shows a MIC assay against MRSA BAA-44 and BAA-1707.
  • FIG. 30 shows a MIC assay against S. epidermidis (ATCC 12228).
  • FIG. 31 shows a MIC assay against S. epidermidis (MRSE 35984).
  • FIG. 32 shows a MIC assay against E. faecium (VRE 700221).
  • FIG. 33 shows co-treatment of MRSA-2 with Tiron and CuSO 4 .
  • FIG. 34 shows co-treatment of S. epidermidis (ATCC 12228) with Tiron and CuSO 4 .
  • FIG. 35 shows co-treatment of E. faecium (VRE 700221) with Tiron and CuSO 4 .
  • novel phenazine derivatives such as compounds of Formulae (I′) (e.g., Formula (I)), (II), and (III), and salts (e.g., pharmaceutically acceptable salts), solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof.
  • the compounds of the invention are expected to be antimicrobial agents and, without wishing to be bound by any particular theory, may act by a microbial warfare strategy (e.g., a reactive oxygen species (ROS)-based competition strategy).
  • a microbial warfare strategy e.g., a reactive oxygen species (ROS)-based competition strategy.
  • the present invention also provides compositions including pharmaceutical compositions, kits, uses, and methods that involve the compounds of the invention and may be useful in preventing and/or treating a microbial infection in a subject, inhibiting the growth and/or reproduction of a microorganism (e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite), killing a microorganism, inhibiting the formation and/or growth of a biofilm, reducing or removing a biofilm, or disinfecting a surface.
  • a microorganism e.g., bacterium, mycobacterium , archaeon, protist,
  • the microorganism is a bacterium.
  • the bacterium is a Gram-positive bacterium (e.g., a species of Staphylococcus or Enterococcus ).
  • the bacterium is a Gram-negative bacterium (e.g., an Acinetobacter species).
  • ROS reactive oxygen species
  • phenazine derivatives such as compounds 301-305 (shown below) are known antimicrobial agents.
  • Pyocyanin (compound 301) is one of the toxins produced by the Gram negative bacterium Pseudomonas aeruginosa . It is thought that Pseudomonas aeruginosa employs a microbial warfare strategy by producing these toxins in competing with other microorganisms (e.g., other bacteria). Pyocyanin is able to oxidize and reduce other molecules (Hassan et al., J.
  • a phenazine derivative may be altered by structurally modifying the phenazine derivative.
  • a known phenazine may be structurally modified to improve its properties, such as antimicrobial activity.
  • the compounds of the invention are improved phenazine derivatives and showed unexpected and superior properties compared to known phenazine derivatives, such as enhanced inhibitory activity against bacteria, e.g., Staphylococcus aureus ( S. aureus ), Staphylococcus epidermidis ( S. epidermidis ), and/or Enterococcus faecium.
  • Staphylococcus aureus is a human pathogen that is notorious for life-threatening drug-resistant infections in hospitals and the community (H. F. Chambers and F. R. DeLeo, Nat. Rev. Microbiol., 2009, 7, 629-641).
  • Staphylococcus aureus MRSA
  • AIDS IDSA Policy Paper d CID 2011:52 (Suppl 5) d S397
  • Staphylococcus epidermidis is also a pathogen of great importance as it is particularly prevalent in persistent microbial infections associated with catheters (I. Uckay, D. Pittet, P. Vaudaux, H. Sax, D. Lew, and F. Waldvogel, Ann. Med., 2009, 41, 109-119).
  • the compounds of the invention may act by a microbial warfare strategy (e.g., an ROS-based competition strategy) similar to the one employed by Pseudomonas aeruginosa .
  • the inventive compounds may be capable of undergoing reduction and oxidation (redox) reactions and forming ROS in, near, or around a microorganism (e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite).
  • a microorganism e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite.
  • An inventive compound may accept a single electron, yielding a relatively stable anion radical, and may readily undergo a redox cycle.
  • a compound of the invention may be reduced by the nicotinamide adenine dinucleotide (NADH + ) in a microorganism and may divert electron flow within the microorganism from the normal cytochrome pathway to an ROS-producing pathway. As a result, the production of ROS, such as O 2 ⁇ and H 2 O 2 , which are toxic to the microorganism, may be increased.
  • NADH + nicotinamide adenine dinucleotide
  • Bacterial biofilms are surface-attached bacterial communities that are encased within a secreted matrix of biomolecules (e.g., extracellular DNA, proteins, polysaccharides) known as the extracellular polymeric substance (EPS).
  • EPS extracellular polymeric substance
  • Bacterial cells within a biofilm take on a completely different physiology than their free-swimming planktonic counterparts and are notorious for being highly resistant to conventional antibiotic treatments and host immune responses (Donlan, R. M. and Costerton, J. W. Clin. Microbiol. Rev. 2002, 15, 167-193).
  • biofilms are present in up to 80% of all bacterial infections.
  • biofilms are notorious for their resistance to conventional antibiotic treatments, and therefore our current arsenal of antibiotics does not include agents that effectively target biofilm machinery or clear established biofilms in a clinical setting.
  • antibiofilm agents would lead to significant breakthroughs in how bacterial infections are treated and would result in the effective treatment of many life-threatening bacterial infections.
  • Bacterial biofilm formation is governed by a signaling process known as quorum sensing, which is used by bacteria to monitor population density and control bacterial virulence (Camilli, A. and Bassler, B. L. Science 2006, 311, 1113-1116; Ng, W.-L. and Bassler, B. L. Annu. Rev. Genet. 2009, 43, 197-222).
  • Quorum sensing is used by free-swimming, individual planktonic bacteria to coordinate the simultaneous attachment and colonization of a surface followed by biofilm formation and maturation.
  • the coordinated surface attachment of bacteria overwhelms immune responses mounted by host organisms, enabling the successful colonization of surfaces (e.g., tissue surfaces) by bacteria.
  • Bacterial biofilms are known to be greater than 1000-fold more resistant to conventional antibiotics when compared to their planktonic counterparts.
  • Therapeutic strategies targeting quorum sensing and/or biofilm formation and dispersion phenotypes have become a promising antibacterial strategy as small molecules capable of inhibiting bacterial biofilm formation via non-growth inhibitory mechanisms or clearing pre-formed bacterial biofilms are of clinical importance.
  • compounds described herein may function by disrupting quorum sensing, leading to inhibitors of biofilm formation and clearing of pre-formed biofilms.
  • the inventive compounds preferably have minimal to no adverse side effects.
  • the compounds exhibit low cytotoxicity against mammalian (e.g., human) cells.
  • the compounds show low hemolysis activity.
  • One aspect of the invention relates to compounds that are believed to be antimicrobial agents.
  • the compounds of the invention are compound of Formula (I′):
  • salts e.g., pharmaceutically acceptable salts
  • solvates hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof, wherein:
  • X is hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl;
  • Y is halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl;
  • R A is hydrogen, halogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR 1 , —N(R 1 ) 2 , —SR 1 , —CN, —SCN, —C( ⁇ NR 1 )R 1 , —C( ⁇ NR 1 )OR 1 , —C( ⁇ NR 1 )N(R 1 ) 2 , —C( ⁇ O)R 1 , —C( ⁇ O)OR 1 , —C( ⁇ O)N(R 1 ) 2 , —NO 2 , —NR C( ⁇ O)R, —NR 1 C
  • R B is hydrogen, halogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR 2 , —N(R 2 ) 2 , —SR 2 , —CN, —SCN, —C( ⁇ NR 2 )R 2 , —C( ⁇ NR 2 )OR 2 , —C( ⁇ NR 2 )N(R 2 ) 2 , —C( ⁇ O)R 2 , —C( ⁇ O)OR 2 , —C( ⁇ O)N(R 2 ) 2 , —NO 2 , —NR 2 C( ⁇ O)R 2 , —
  • R A and R B are joined to form a substituted or unsubstituted phenyl ring;
  • At least one of X and Y is halogen
  • the compounds of the invention are compounds of Formula (I):
  • salts e.g., pharmaceutically acceptable salts
  • solvates hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof, wherein:
  • X is hydrogen or halogen
  • Y is halogen
  • R A is hydrogen, halogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR 1 , —N(R 1 ) 2 , —SR 1 , —CN, —SCN, —C( ⁇ NR 1 )R 1 , —C( ⁇ NR 1 )OR 1 , —C( ⁇ NR 1 )N(R 1 ) 2 , —C( ⁇ O)R 1 , —C( ⁇ O)OR 1 , —C( ⁇ O)N(R 1 ) 2 , —NO 2 , —NR C( ⁇ O)R, —NR 1 C
  • R B is hydrogen, halogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR 2 , —N(R 2 ) 2 , —SR 2 , —CN, —SCN, —C( ⁇ NR 2 )R 2 , —C( ⁇ NR 2 )OR 2 , —C( ⁇ NR 2 )N(R 2 ) 2 , —C( ⁇ O)R 2 , —C( ⁇ O)OR 2 , —C( ⁇ O)N(R 2 ) 2 , —NO 2 , —NR 2 C( ⁇ O)R 2 , —
  • R A and R B are joined to form a substituted or unsubstituted phenyl ring; and provided that the compound is not of the formula:
  • Formula (I′) (e.g., Formula (I)) includes substituent X on the phenazinyl ring.
  • X is hydrogen.
  • X is halogen.
  • X is F.
  • X is Cl.
  • X is Br.
  • X is I.
  • X is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl).
  • X is Me.
  • X is substituted methyl (e.g., —CH 2 F, —CHF 2 , —CF 3 , or Bn).
  • X is Et, substituted ethyl (e.g., fluorinated ethyl (e.g., perfluoroethyl)), Pr, substituted propyl (e.g., fluorinated propyl (e.g., perfluoropropyl)), Bu, substituted butyl (e.g., fluorinated butyl (e.g., perfluorobutyl)), unsubstituted pentyl, substituted pentyl (e.g., fluorinated pentyl (e.g., perfluoropentyl)), unsubstituted hexyl, or substituted hexyl (e.g., fluorinated hexyl (e.g., perfluorohexyl)).
  • substituted ethyl e.g., fluorinated ethyl (e.g., perfluoroethyl)
  • Pr substituted
  • X is substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl). In certain embodiments, X is substituted or unsubstituted vinyl. In certain embodiments, X is unsubstituted allyl. In certain embodiments, X is substituted allyl. In certain embodiments, X is substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl).
  • Formula (I′) also includes substituent Y on the phenazinyl ring.
  • Y is halogen.
  • Y is F.
  • Y is C 1 .
  • Y is Br.
  • Y is I.
  • Y is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl).
  • Y is Me.
  • Y is substituted methyl (e.g., —CH 2 F, —CHF 2 , —CF 3 , or Bn).
  • Y is Et, substituted ethyl (e.g., fluorinated ethyl (e.g., perfluoroethyl)), Pr, substituted propyl (e.g., fluorinated propyl (e.g., perfluoropropyl)), Bu, substituted butyl (e.g., fluorinated butyl (e.g., perfluorobutyl)), unsubstituted pentyl, substituted pentyl (e.g., fluorinated pentyl (e.g., perfluoropentyl)), unsubstituted hexyl, or substituted hexyl (e.g., fluorinated hexyl (e.g., perfluorohexyl)).
  • substituted ethyl e.g., fluorinated ethyl (e.g., perfluoroethyl)
  • Pr substituted
  • Y is n-Bu. In certain embodiments, Y is substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl). In certain embodiments, Y is substituted or unsubstituted vinyl. In certain embodiments, Y is unsubstituted allyl. In certain embodiments, Y is substituted allyl. In certain embodiments, Y is substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl).
  • X is hydrogen; and Y is F. In certain embodiments, X is hydrogen; and Y is Cl. In certain embodiments, X is hydrogen; and Y is Br. In certain embodiments, X is hydrogen; and Y is I. In certain embodiments, X is Cl; and Y is F. In certain embodiments, both X and Y are Cl. In certain embodiments, X is Cl; and Y is Br. In certain embodiments, X is Cl; and Y is I. In certain embodiments, X is Br; and Y is F. In certain embodiments, X is Br; and Y is Cl. In certain embodiments, both X and Y are Br. In certain embodiments, X is Br; and Y is Br. In certain embodiments, X is Br; and Y is I.
  • X is I; and Y is F. In certain embodiments, X is I; and Y is Cl. In certain embodiments, X is I; and Y is Br. In certain embodiments, both X and Y are I.
  • X is halogen; and Y is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl). In certain embodiments, X is halogen; and Y is substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl).
  • X is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl); and Y is halogen.
  • X is substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl); and Y is halogen.
  • X is halogen; and X and Y are the same.
  • X is halogen; and X and Y are not the same.
  • at least one of X and Y is halogen.
  • each X and Y is halogen.
  • Formula (I′) also includes substituent R A on the phenazinyl ring.
  • R A is hydrogen. In certain embodiments, R A is not hydrogen. In certain embodiments, R A is halogen. In certain embodiments, R A is F. In certain embodiments, R A is C 1 . In certain embodiments, R A is Br. In certain embodiments, R A is I. In certain embodiments, R A is substituted or unsubstituted alkyl. In certain embodiments, R A is substituted or unsubstituted C 1-6 alkyl. In certain embodiments, R A is Me.
  • R A is substituted methyl (e.g., —CH 2 F, —CHF 2 , —CF 3 , or Bn).
  • R A is Et, substituted ethyl (e.g., fluorinated ethyl (e.g., perfluoroethyl)), Pr, substituted propyl (e.g., fluorinated propyl (e.g., perfluoropropyl)), Bu, or substituted butyl (e.g., fluorinated butyl (e.g., perfluorobutyl)).
  • R A is substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl). In certain embodiments, R A is substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl). In certain embodiments, R A is substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system).
  • R A is substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, or substituted or unsubstituted cyclohexyl.
  • R A is substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur).
  • R A is substituted or unsubstituted oxetanyl, substituted or unsubstituted tetrahydrofuranyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted morpholinyl, or substituted or unsubstituted piperazinyl.
  • R A is substituted or unsubstituted aryl (e.g., substituted or unsubstituted, 6- to 10-membered aryl).
  • R A is unsubstituted phenyl.
  • R A is substituted phenyl. In certain embodiments, R A is substituted or unsubstituted naphthyl. In certain embodiments, R A is substituted or unsubstituted heteroaryl. In certain embodiments, R A is substituted or unsubstituted, 5- to 6-membered, monocyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In certain embodiments, R A is substituted or unsubstituted, 9- to 10-membered, bicyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur.
  • R A is —OR 1 (e.g., —OH, —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe, —OCF 3 , —OEt, —OPr, —OBu, or —OBn), or —O(substituted or unsubstituted phenyl) (e.g., —OPh)).
  • R A is —OMe.
  • R A is —SR 1 (e.g., —SH, —S(substituted or unsubstituted C 1-6 alkyl) (e.g., —SMe, —SEt, —SPr, —SBu, or —SBn), or —S(substituted or unsubstituted phenyl) (e.g., —SPh)).
  • SR 1 e.g., —SH, —S(substituted or unsubstituted C 1-6 alkyl) (e.g., —SMe, —SEt, —SPr, —SBu, or —SBn), or —S(substituted or unsubstituted phenyl) (e.g., —SPh)).
  • R A is —N(R 1 ) 2 (e.g., —NH 2 , —NH(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHMe), or —N(substituted or unsubstituted C 1-6 alkyl)-(substituted or unsubstituted C 1-6 alkyl) (e.g., —NMe 2 )).
  • R A is —CN or —SCN.
  • R A is —NO 2 .
  • R A is —C( ⁇ NR 1 )R 1 , —C( ⁇ NR 1 )OR 1 , or —C( ⁇ NR 1 )N(R 1 ) 2 .
  • R A is —C( ⁇ O)R 1 (e.g., —C( ⁇ O)(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)Me) or —C( ⁇ O)(substituted or unsubstituted phenyl)).
  • R A is —C( ⁇ O)OR 1 (e.g., —C( ⁇ O)OH, —C( ⁇ O)O(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)OMe), or —C( ⁇ O)O(substituted or unsubstituted phenyl)).
  • R A is —C( ⁇ O)OR 1 (e.g., —C( ⁇ O)OH, —C( ⁇ O)O(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)OMe), or —C( ⁇ O)O(substituted or unsubstituted phenyl)).
  • R A is —C( ⁇ O)N(R 1 ) 2 (e.g., —C( ⁇ O)NH 2 , —C( ⁇ O)NH(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)NHMe), —C( ⁇ O)NH(substituted or unsubstituted phenyl), —C( ⁇ O)N(substituted or unsubstituted alkyl)-(substituted or unsubstituted alkyl), or —C( ⁇ O)N(substituted or unsubstituted phenyl)-(substituted or unsubstituted alkyl)).
  • R A is —C( ⁇ O)N(R 1 ) 2 (e.g., —C( ⁇ O)NH 2 , —C( ⁇ O)NH(substituted or unsubstituted alkyl) (e
  • R A is —NR 1 C( ⁇ O)R 1 (e.g., —NHC( ⁇ O)(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHC( ⁇ O)Me) or —NHC( ⁇ O)(substituted or unsubstituted phenyl)).
  • R A is —NR 1 C( ⁇ O)OR 1 .
  • R A is —NR 1 C( ⁇ O)N(R 1 ) 2 (e.g., —NHC( ⁇ O)NH 2 , —NHC( ⁇ O)NH(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHC( ⁇ O)NHMe)).
  • R A is —OC( ⁇ O)R 1 (e.g., —OC( ⁇ O)(substituted or unsubstituted alkyl) or —OC( ⁇ O)(substituted or unsubstituted phenyl)), —OC( ⁇ O)OR 1 (e.g., —OC( ⁇ O)O(substituted or unsubstituted alkyl) or —OC( ⁇ O)O(substituted or unsubstituted phenyl)), or —OC( ⁇ O)N(R 1 ) 2 (e.g., —OC( ⁇ O)NH 2 , —OC( ⁇ O)NH(substituted or unsubstituted alkyl), —OC( ⁇ O)NH(substituted or unsubstituted phenyl), —OC( ⁇ O)N(substituted or unsubstituted alkyl)-(substit
  • Formula (I′) may include one or more instances of substituent R 1 .
  • Formula (I′) e.g., Formula (I)
  • any two instances of R 1 may be the same or different from each other.
  • at least one instance of R 1 is H.
  • each instance of R 1 is H.
  • at least one instance of R 1 is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)).
  • R 1 is substituted or unsubstituted acyl, substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl), substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl), substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system), substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or
  • Formula (I′) also includes substituent R B on the phenazinyl ring.
  • R B is hydrogen. In certain embodiments, R B is not hydrogen. In certain embodiments, R B is halogen. In certain embodiments, R B is F. In certain embodiments, R B is Cl. In certain embodiments, R B is Br. In certain embodiments, R B is I. In certain embodiments, R B is substituted or unsubstituted alkyl. In certain embodiments, R B is substituted or unsubstituted C 1-6 alkyl. In certain embodiments, R B is Me.
  • R B is substituted methyl (e.g., —CH 2 F, —CHF 2 , —CF 3 , or Bn).
  • R B is Et, substituted ethyl (e.g., fluorinated ethyl (e.g., perfluoroethyl)), Pr, substituted propyl (e.g., fluorinated propyl (e.g., perfluoropropyl)), Bu, or substituted butyl (e.g., fluorinated butyl (e.g., perfluorobutyl)).
  • R B is substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl). In certain embodiments, R B is substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl). In certain embodiments, R B is substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system).
  • R B is substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, or substituted or unsubstituted cyclohexyl.
  • R B is substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur).
  • R B is substituted or unsubstituted oxetanyl, substituted or unsubstituted tetrahydrofuranyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted morpholinyl, or substituted or unsubstituted piperazinyl.
  • R B is substituted or unsubstituted aryl (e.g., substituted or unsubstituted, 6- to 10-membered aryl).
  • R B is unsubstituted phenyl.
  • R B is substituted phenyl. In certain embodiments, R B is substituted or unsubstituted naphthyl. In certain embodiments, R B is substituted or unsubstituted heteroaryl. In certain embodiments, R B is substituted or unsubstituted, 5- to 6-membered, monocyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In certain embodiments, R B is substituted or unsubstituted, 9- to 10-membered, bicyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur.
  • R B is —OR 2 (e.g., —OH, —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe, —OCF 3 , —OEt, —OPr, —OBu, or —OBn), or —O(substituted or unsubstituted phenyl) (e.g., —OPh)).
  • R B is —OMe.
  • R B is —SR 2 (e.g., —SH, —S(substituted or unsubstituted C 1-6 alkyl) (e.g., —SMe, —SEt, —SPr, —SBu, or —SBn), or —S(substituted or unsubstituted phenyl) (e.g., —SPh)).
  • SR 2 e.g., —SH, —S(substituted or unsubstituted C 1-6 alkyl) (e.g., —SMe, —SEt, —SPr, —SBu, or —SBn), or —S(substituted or unsubstituted phenyl) (e.g., —SPh)).
  • R B is —N(R 2 ) 2 (e.g., —NH 2 , —NH(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHMe), or —N(substituted or unsubstituted C 1-6 alkyl)-(substituted or unsubstituted C 1-6 alkyl) (e.g., —NMe 2 )).
  • R B is —CN or —SCN.
  • R B is —NO 2 .
  • R B is —C( ⁇ NR 2 )R 2 , —C( ⁇ NR 2 )OR 2 , or —C( ⁇ NR 2 )N(R 2 ) 2 .
  • R B is —C( ⁇ O)R 2 (e.g., —C( ⁇ O)(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)Me) or —C( ⁇ O)(substituted or unsubstituted phenyl)).
  • R B is —C( ⁇ O)OR 2 (e.g., —C( ⁇ O)OH, —C( ⁇ O)O(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)OMe), or —C( ⁇ O)O(substituted or unsubstituted phenyl)).
  • R B is —C( ⁇ O)OR 2 (e.g., —C( ⁇ O)OH, —C( ⁇ O)O(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)OMe), or —C( ⁇ O)O(substituted or unsubstituted phenyl)).
  • R B is —C( ⁇ O)N(R 2 ) 2 (e.g., —C( ⁇ O)NH 2 , —C( ⁇ O)NH(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)NHMe), —C( ⁇ O)NH(substituted or unsubstituted phenyl), —C( ⁇ O)N(substituted or unsubstituted alkyl)-(substituted or unsubstituted alkyl), or —C( ⁇ O)N(substituted or unsubstituted phenyl)-(substituted or unsubstituted alkyl)).
  • R B is —C( ⁇ O)N(R 2 ) 2 (e.g., —C( ⁇ O)NH 2 , —C( ⁇ O)NH(substituted or unsubstituted alkyl) (e
  • R B is —NR 2 C( ⁇ O)R 2 (e.g., —NHC( ⁇ O)(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHC( ⁇ O)Me) or —NHC( ⁇ O)(substituted or unsubstituted phenyl)).
  • R B is —NR 2 C( ⁇ O)OR 2 .
  • R B is —NR 2 C( ⁇ O)N(R 2 ) 2 (e.g., —NHC( ⁇ O)NH 2 , —NHC( ⁇ O)NH(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHC( ⁇ O)NHMe)).
  • R B is —OC( ⁇ O)R 2 (e.g., —OC( ⁇ O)(substituted or unsubstituted alkyl) or —OC( ⁇ O)(substituted or unsubstituted phenyl)), —OC( ⁇ O)OR 2 (e.g., —OC( ⁇ O)O(substituted or unsubstituted alkyl) or —OC( ⁇ O)O(substituted or unsubstituted phenyl)), or —OC( ⁇ O)N(R 2 ) 2 (e.g., —OC( ⁇ O)NH 2 , —OC( ⁇ O)NH(substituted or unsubstituted alkyl), —OC( ⁇ O)NH(substituted or unsubstituted phenyl), —OC( ⁇ O)N(substituted or unsubstituted alkyl)-(substit
  • Formula (I′) may include one or more instances of substituent R 2 .
  • Formula (I′) e.g., Formula (I)
  • any two instances of R 2 may be the same or different from each other.
  • at least one instance of R 2 is H.
  • each instance of R 2 is H.
  • at least one instance of R 2 is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)).
  • R 2 is substituted or unsubstituted acyl, substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl), substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl), substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system), substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or
  • both R A and R B are H.
  • each of R A and R B is independently hydrogen or halogen.
  • each of R A and R B is independently hydrogen, halogen, or substituted or unsubstituted C 1-6 alkyl (e.g., Me).
  • each of R A and R B is independently hydrogen, halogen, substituted or unsubstituted C 1-6 alkyl (e.g., Me), or —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe).
  • R A is H; and R B is halogen (e.g., F, Cl, Br, or I).
  • R A is H; and R B is substituted or unsubstituted C 1-6 alkyl (e.g., Me).
  • R A is halogen (e.g., F, Cl, Br, and I); and R B is H.
  • R A is substituted or unsubstituted C 1-6 alkyl; and R B is H.
  • both R A and R B are halogen.
  • both R A and R B are halogen; and R A and R B are the same.
  • both R A and R B are halogen; and R A and R B are not the same.
  • each of R A and R B is independently Cl, Br, or I.
  • both R A and R B are C 1 . In certain embodiments, R A is Cl; and R B is Br. In certain embodiments, R A is Cl; and R B is I. In certain embodiments, R A is Br; and R B is C 1 . In certain embodiments, both R A and R B are Br. In certain embodiments, R A is Br; and R B is I. In certain embodiments, R A is I; and R B is C 1 . In certain embodiments, R A is I; and R B is Br. In certain embodiments, both R A and R B are I. In certain embodiments, both R A and R B are substituted or unsubstituted C 1-6 alkyl (e.g., Me).
  • R A is substituted or unsubstituted C 1-6 alkyl (e.g., Me); and R B is halogen (e.g., Cl, Br, or I). In certain embodiments, R A is halogen (e.g., Cl, Br, or I); and R B is substituted or unsubstituted C 1-6 alkyl (e.g., Me).
  • R A is —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe); and R B is hydrogen, halogen, substituted or unsubstituted C 1-6 alkyl (e.g., Me), or —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe).
  • R A is hydrogen, halogen, substituted or unsubstituted C 1-6 alkyl (e.g., Me), or —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe); and R B is —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe).
  • a compound of the invention is not of the formula:
  • a compound of the invention is not of the formula:
  • At least one of R A and R B is not hydrogen. In certain embodiments, both R A and R B are not hydrogen.
  • a compound of the invention is not of the formula:
  • each of R A and R B is independently halogen or unsubstituted C 1-6 alkyl.
  • a compound of the invention is not of the formula:
  • each of R A and R B is independently Cl, Br, or Me.
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • X is halogen
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A is not hydrogen (e.g., wherein R A is halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R B is not hydrogen (e.g., wherein R B is halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • R A and R B are not hydrogen (e.g., wherein R A and R B is independently halogen or substituted or unsubstituted C 1-6 alkyl).
  • the compound of Formula (I′) (e.g., Formula (I)) is of the formula:
  • Exemplary compounds of Formula (I′) include:
  • Exemplary compounds of Formula (I′) also include:
  • Exemplary compounds of Formula (I′) also include:
  • Exemplary compounds of Formula (I′) also include:
  • Exemplary compounds of Formula (I′) also include:
  • Exemplary compounds of Formula (I′) also include:
  • Exemplary compounds of Formula (I′) also include:
  • Exemplary compounds of Formula (I′) also include:
  • Exemplary compounds of Formula (I′) also include:
  • Exemplary compounds of Formula (I′) also include:
  • the compounds of the invention are compounds of Formula (II):
  • W is hydrogen or halogen
  • Z is halogen
  • R C is hydrogen, halogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR 3 , —N(R 3 ) 2 , —SR 3 , —CN, —SCN, —C( ⁇ NR 3 )R 3 , —C( ⁇ NR 3 )OR 3 , —C( ⁇ NR 3 )N(R 3 ) 2 , —C( ⁇ O)R 3 , —C( ⁇ O)OR 3 , —C( ⁇ O)N(R 3 ) 2 , —NO 2 , —NR 3 C( ⁇ O)R 3 , —
  • R D is hydrogen, halogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR 4 , —N(R 4 ) 2 , —SR 4 , —CN, —SCN, —C( ⁇ NR 4 )R 4 , —C( ⁇ NR 4 )OR 4 , —C( ⁇ NR 4 )N(R 4 ) 2 , —C( ⁇ O)R 4 , —C( ⁇ O)OR 4 , —C( ⁇ O)N(R 4 ) 2 , —NO 2 , —NR 4 C( ⁇ O)R 4 , —
  • the compounds of the invention are compounds of Formula of Formula (III):
  • W is hydrogen or halogen
  • Z is halogen
  • R C is hydrogen, halogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR 3 , —N(R 3 ) 2 , —SR 3 , —CN, —SCN, —C( ⁇ NR 3 )R 3 , —C( ⁇ NR 3 )OR 3 , —C( ⁇ NR 3 )N(R 3 ) 2 , —C( ⁇ O)R 3 , —C( ⁇ O)OR 3 , —C( ⁇ O)N(R 3 ) 2 , —NO 2 , —NR 3 C( ⁇ O)R 3 , —
  • R D is hydrogen, halogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR 4 , —N(R 4 ) 2 , —SR 4 , —CN, —SCN, —C( ⁇ NR 4 )R 4 , —C( ⁇ NR 4 )OR 4 , —C( ⁇ NR 4 )N(R 4 ) 2 , —C( ⁇ O)R 4 , —C( ⁇ O)OR 4 , —C( ⁇ O)N(R 4 ) 2 , —NO 2 , —NR 4 C( ⁇ O)R 4 , —
  • Formula (II) or (III) includes substituent W on the quinoxalinyl ring.
  • W is hydrogen.
  • W is halogen.
  • W is F.
  • W is C 1 .
  • W is Br.
  • W is I.
  • Formula (II) or (III) also includes substituent Z on the quinoxalinyl ring.
  • Z is F.
  • Z is C 1 .
  • Z is Br.
  • Z is I.
  • W is Cl; and Z is F. In certain embodiments, both W and Z and C 1 . In certain embodiments, W is Cl; and Z is Br. In certain embodiments, W is Cl; and Z is I. In certain embodiments, W is Br; and Z is F. In certain embodiments, W is Br; and Z is C 1 . In certain embodiments, both W and Z are Br. In certain embodiments, W is Br; and Z is I. In certain embodiments, W is I; and Z is F. In certain embodiments, W is I; and Z is C 1 . In certain embodiments, W is I; and Z is Br. In certain embodiments, both W and Z are I. In certain embodiments, W and Z are the same. In certain embodiments, W and Z are not the same.
  • Formula (II) or (III) also includes substituent R C on the quinoxalinyl ring.
  • R C is hydrogen. In certain embodiments, R C is not hydrogen. In certain embodiments, R C is halogen. In certain embodiments, R C is F. In certain embodiments, R C is C 1 . In certain embodiments, R C is Br. In certain embodiments, R C is I. In certain embodiments, R C is substituted or unsubstituted alkyl. In certain embodiments, R C is substituted or unsubstituted C 1-6 alkyl. In certain embodiments, R cc is Me.
  • R C is substituted methyl (e.g., —CH 2 F, —CHF 2 , —CF 3 , or Bn).
  • R C is Et, substituted ethyl (e.g., fluorinated ethyl (e.g., perfluoroethyl)), Pr, substituted propyl (e.g., fluorinated propyl (e.g., perfluoropropyl)), Bu, or substituted butyl (e.g., fluorinated butyl (e.g., perfluorobutyl)).
  • R C is substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl). In certain embodiments, R C is substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl). In certain embodiments, R C is substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system).
  • R C is substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, or substituted or unsubstituted cyclohexyl.
  • R C is substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur).
  • R C is substituted or unsubstituted oxetanyl, substituted or unsubstituted tetrahydrofuranyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted morpholinyl, or substituted or unsubstituted piperazinyl.
  • R C is substituted or unsubstituted aryl (e.g., substituted or unsubstituted, 6- to 10-membered aryl).
  • R C is unsubstituted phenyl.
  • R C is substituted phenyl. In certain embodiments, R C is substituted or unsubstituted naphthyl. In certain embodiments, R C is substituted or unsubstituted heteroaryl. In certain embodiments, R C is substituted or unsubstituted, 5- to 6-membered, monocyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In certain embodiments, R C is substituted or unsubstituted, 9- to 10-membered, bicyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur.
  • R C is —OR 3 (e.g., —OH, —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe, —OCF 3 , —OEt, —OPr, —OBu, or —OBn), or —O(substituted or unsubstituted phenyl) (e.g., —OPh)).
  • OR 3 e.g., —OH, —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe, —OCF 3 , —OEt, —OPr, —OBu, or —OBn), or —O(substituted or unsubstituted phenyl) (e.g., —OPh)).
  • R C is —SR 3 (e.g., —SH, —S(substituted or unsubstituted C 1-6 alkyl) (e.g., —SMe, —SEt, —SPr, —SBu, or —SBn), or —S(substituted or unsubstituted phenyl) (e.g., —SPh)).
  • SR 3 e.g., —SH, —S(substituted or unsubstituted C 1-6 alkyl) (e.g., —SMe, —SEt, —SPr, —SBu, or —SBn), or —S(substituted or unsubstituted phenyl) (e.g., —SPh)).
  • R C is —N(R 3 ) 2 (e.g., —NH 2 , —NH(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHMe), or —N(substituted or unsubstituted C 1-6 alkyl)-(substituted or unsubstituted C 1-6 alkyl) (e.g., —NMe 2 )).
  • R C is —CN or —SCN.
  • R C is —NO 2 .
  • R C is —C( ⁇ NR 3 )R 3 , —C( ⁇ NR 3 )OR 3 , or —C( ⁇ NR 3 )N(R 3 ) 2 .
  • R C is —C( ⁇ O)R 3 (e.g., —C( ⁇ O)(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)Me) or —C( ⁇ O)(substituted or unsubstituted phenyl)).
  • R C is —C( ⁇ O)OR 3 (e.g., —C( ⁇ O)OH, —C( ⁇ O)O(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)OMe), or —C( ⁇ O)O(substituted or unsubstituted phenyl)).
  • R C is —C( ⁇ O)OR 3 (e.g., —C( ⁇ O)OH, —C( ⁇ O)O(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)OMe), or —C( ⁇ O)O(substituted or unsubstituted phenyl)).
  • R C is —C( ⁇ O)N(R 3 ) 2 (e.g., —C( ⁇ O)NH 2 , —C( ⁇ O)NH(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)NHMe), —C( ⁇ O)NH(substituted or unsubstituted phenyl), —C( ⁇ O)N(substituted or unsubstituted alkyl)-(substituted or unsubstituted alkyl), or —C( ⁇ O)N(substituted or unsubstituted phenyl)-(substituted or unsubstituted alkyl)).
  • R C is —C( ⁇ O)N(R 3 ) 2 (e.g., —C( ⁇ O)NH 2 , —C( ⁇ O)NH(substituted or unsubstituted alkyl) (e
  • R C is —NR 3 C( ⁇ O)R 3 (e.g., —NHC( ⁇ O)(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHC( ⁇ O)Me) or —NHC( ⁇ O)(substituted or unsubstituted phenyl)).
  • R C is —NR 3 C( ⁇ O)OR 3 .
  • R C is —NR 3 C( ⁇ O)N(R 3 ) 2 (e.g., —NHC( ⁇ O)NH 2 , —NHC( ⁇ O)NH(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHC( ⁇ O)NHMe)).
  • R C is —OC( ⁇ O)R 3 (e.g., —OC( ⁇ O)(substituted or unsubstituted alkyl) or —OC( ⁇ O)(substituted or unsubstituted phenyl)), —OC( ⁇ O)OR 3 (e.g., —OC( ⁇ O)O(substituted or unsubstituted alkyl) or —OC( ⁇ O)O(substituted or unsubstituted phenyl)), or —OC( ⁇ O)N(R 3 ) 2 (e.g., —OC( ⁇ O)NH 2 , —OC( ⁇ O)NH(substituted or unsubstituted alkyl), —OC( ⁇ O)NH(substituted or unsubstituted phenyl), —OC( ⁇ O)N(substituted or unsubstituted alkyl)-(substit
  • Formula (II) or (III) may include one or more instances of substituent R 3 .
  • any two instances of R 3 may be the same or different from each other.
  • at least one instance of R 3 is H.
  • each instance of R 3 is H.
  • at least one instance of R 3 is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)).
  • R 3 is substituted or unsubstituted acyl, substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl), substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl), substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system), substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or
  • Formula (II) or (III) also includes substituent R D on the quinoxalinyl ring.
  • R D is hydrogen. In certain embodiments, R D is not hydrogen. In certain embodiments, R D is halogen. In certain embodiments, R D is F. In certain embodiments, R D is C 1 . In certain embodiments, R D is Br. In certain embodiments, R D is I. In certain embodiments, R D is substituted or unsubstituted alkyl. In certain embodiments, R D is substituted or unsubstituted C 1-6 alkyl. In certain embodiments, R D is Me.
  • R D is substituted methyl (e.g., —CH 2 F, —CHF 2 , —CF 3 , or Bn).
  • R D is Et, substituted ethyl (e.g., fluorinated ethyl (e.g., perfluoroethyl)), Pr, substituted propyl (e.g., fluorinated propyl (e.g., perfluoropropyl)), Bu, or substituted butyl (e.g., fluorinated butyl (e.g., perfluorobutyl)).
  • R D is substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl). In certain embodiments, R D is substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl). In certain embodiments, R D is substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system).
  • R D is substituted or unsubstituted cyclopropyl, substituted or unsubstituted cyclobutyl, substituted or unsubstituted cyclopentyl, or substituted or unsubstituted cyclohexyl.
  • R D is substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or sulfur).
  • R D is substituted or unsubstituted oxetanyl, substituted or unsubstituted tetrahydrofuranyl, substituted or unsubstituted pyrrolidinyl, substituted or unsubstituted tetrahydropyranyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted morpholinyl, or substituted or unsubstituted piperazinyl.
  • R D is substituted or unsubstituted aryl (e.g., substituted or unsubstituted, 6- to 10-membered aryl).
  • R D is unsubstituted phenyl.
  • R D is substituted phenyl. In certain embodiments, R D is substituted or unsubstituted naphthyl. In certain embodiments, R D is substituted or unsubstituted heteroaryl. In certain embodiments, R D is substituted or unsubstituted, 5- to 6-membered, monocyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In certain embodiments, R D is substituted or unsubstituted, 9- to 10-membered, bicyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur.
  • R D is —OR 4 (e.g., —OH, —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe, —OCF 3 , —OEt, —OPr, —OBu, or —OBn), or —O(substituted or unsubstituted phenyl) (e.g., —OPh)).
  • OR 4 e.g., —OH, —O(substituted or unsubstituted C 1-6 alkyl) (e.g., —OMe, —OCF 3 , —OEt, —OPr, —OBu, or —OBn), or —O(substituted or unsubstituted phenyl) (e.g., —OPh)).
  • R D is —SR 4 (e.g., —SH, —S(substituted or unsubstituted C 1-6 alkyl) (e.g., —SMe, —SEt, —SPr, —SBu, or —SBn), or —S(substituted or unsubstituted phenyl) (e.g., —SPh)).
  • SR 4 e.g., —SH, —S(substituted or unsubstituted C 1-6 alkyl) (e.g., —SMe, —SEt, —SPr, —SBu, or —SBn), or —S(substituted or unsubstituted phenyl) (e.g., —SPh)).
  • R D is —N(R 4 ) 2 (e.g., —NH 2 , —NH(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHMe), or —N(substituted or unsubstituted C 1-6 alkyl)-(substituted or unsubstituted C 1-6 alkyl) (e.g., —NMe 2 )).
  • R D is —CN or —SCN.
  • R D is —NO 2 .
  • R D is —C( ⁇ NR 4 )R 4 , —C( ⁇ NR 4 )OR 4 , or —C( ⁇ NR 4 )N(R 4 ) 2 .
  • R D is —C( ⁇ O)R 4 (e.g., —C( ⁇ O)(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)Me) or —C( ⁇ O)(substituted or unsubstituted phenyl)).
  • R D is —C( ⁇ O)OR 4 (e.g., —C( ⁇ O)OH, —C( ⁇ O)O(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)OMe), or —C( ⁇ O)O(substituted or unsubstituted phenyl)).
  • R D is —C( ⁇ O)OR 4 (e.g., —C( ⁇ O)OH, —C( ⁇ O)O(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)OMe), or —C( ⁇ O)O(substituted or unsubstituted phenyl)).
  • R D is —C( ⁇ O)N(R 4 ) 2 (e.g., —C( ⁇ O)NH 2 , —C( ⁇ O)NH(substituted or unsubstituted alkyl) (e.g., —C( ⁇ O)NHMe), —C( ⁇ O)NH(substituted or unsubstituted phenyl), —C( ⁇ O)N(substituted or unsubstituted alkyl)-(substituted or unsubstituted alkyl), or —C( ⁇ O)N(substituted or unsubstituted phenyl)-(substituted or unsubstituted alkyl)).
  • R D is —C( ⁇ O)N(R 4 ) 2 (e.g., —C( ⁇ O)NH 2 , —C( ⁇ O)NH(substituted or unsubstituted alkyl) (e
  • R D is —NR 4 C( ⁇ O)R 4 (e.g., —NHC( ⁇ O)(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHC( ⁇ O)Me) or —NHC( ⁇ O)(substituted or unsubstituted phenyl)).
  • R D is —NR 4 C( ⁇ O)OR 4 .
  • R D is —NR 4 C( ⁇ O)N(R 4 ) 2 (e.g., —NHC( ⁇ O)NH 2 , —NHC( ⁇ O)NH(substituted or unsubstituted C 1-6 alkyl) (e.g., —NHC( ⁇ O)NHMe)).
  • R D is —OC( ⁇ O)R 4 (e.g., —OC( ⁇ O)(substituted or unsubstituted alkyl) or —OC( ⁇ O)(substituted or unsubstituted phenyl)), —OC( ⁇ O)OR 4 (e.g., —OC( ⁇ O)O(substituted or unsubstituted alkyl) or —OC( ⁇ O)O(substituted or unsubstituted phenyl)), or —OC( ⁇ O)N(R 4 ) 2 (e.g., —OC( ⁇ O)NH 2 , —OC( ⁇ O)NH(substituted or unsubstituted alkyl), —OC( ⁇ O)NH(substituted or unsubstituted phenyl), —OC( ⁇ O)N(substituted or unsubstituted alkyl)-(substit
  • Formula (II) or (III) may include one or more instances of substituent R 4 .
  • any two instances of R 4 may be the same or different from each other.
  • at least one instance of R 4 is H.
  • each instance of R 4 is H.
  • at least one instance of R 4 is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)).
  • R 4 is substituted or unsubstituted acyl, substituted or unsubstituted alkenyl (e.g., substituted or unsubstituted C 2-6 alkenyl), substituted or unsubstituted alkynyl (e.g., substituted or unsubstituted C 2-6 alkynyl), substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system), substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic heterocyclyl comprising zero, one, or two double bonds in the heterocyclic ring system, wherein one, two, or three atoms in the heterocyclic ring system are independently nitrogen, oxygen, or
  • R C is hydrogen, and R D is not hydrogen.
  • R D is hydrogen, and R C is not hydrogen.
  • each one of R C and R D is not hydrogen.
  • both R C and R D are H.
  • both R C and R D are halogen (e.g., Cl, Br, or I).
  • both R C and R D are substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)).
  • each of R C and R D is independently hydrogen or substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)). In certain embodiments, each of R C and R D is independently hydrogen, halogen (e.g., Cl, Br, or I), or substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)).
  • halogen e.g., Cl, Br, or I
  • each of R C and R D is independently halogen (e.g., Cl, Br, or I) or substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)).
  • halogen e.g., Cl, Br, or I
  • substituted or unsubstituted alkyl e.g., substituted or unsubstituted C 1-6 alkyl (e.g., Me)
  • the compound of the invention is not of the formula:
  • R C and R D is not unsubstituted C 1-6 alkyl (e.g., Me). In certain embodiments, only one of R C and R D is not unsubstituted C 1-6 alkyl (e.g., Me). In certain embodiments, at least one of W and Z is not Br.
  • Exemplary compounds of Formula (II) include:
  • Exemplary compounds of Formula (II) further include:
  • Exemplary compounds of Formula (III) include:
  • the compounds of the invention are the compounds described herein, and salts (e.g., pharmaceutically acceptable salts), solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof.
  • the compounds of the invention are the compounds described herein, and pharmaceutically acceptable salts thereof.
  • the compounds of the invention are the compounds described herein, and pharmaceutically acceptable salts thereof.
  • the compounds of the invention are the compounds of Formula (I′), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof.
  • the compounds of the invention are the compounds of Formula (I′), and salts (e.g., pharmaceutically acceptable salts) thereof.
  • the compounds of the invention are the compounds of Formula (I), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof.
  • the compounds of the invention are the compounds of Formula (I), and salts (e.g., pharmaceutically acceptable salts) thereof.
  • the compounds of the invention are the compounds of Formula (II), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof.
  • the compounds of the invention are the compounds of Formula (II), and salts (e.g., pharmaceutically acceptable salts) thereof.
  • the compounds of the invention are the compounds of Formula (III), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof.
  • the compounds of the invention are the compounds of Formula (III), and salts (e.g., pharmaceutically acceptable salts) thereof.
  • the compounds of the invention are substantially pure. In certain embodiments, a compound of the invention is at least about 90%, at least about 95%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% free of impurities.
  • the compounds of the invention have been found to be antimicrobial agents (e.g., antibacterial agents).
  • the compounds of the invention may be redox-active and may generate reactive oxygen species (ROS).
  • ROS reactive oxygen species
  • the inventive compounds may thus act as microbial warfare agents and inhibit the growth and/or reproduction of or kill a microorganism (e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite) by oxidizing and/or reducing molecules (e.g., catalase, cytokine, nicotinamide adenine dinucleotide phosphate (NADPH), and nicotinamide adenine dinucleotide phosphate (NADP + )) in, near, or around the microorganism.
  • a microorganism e.g., bacterium, mycobacterium , archaeon, protist, fungus, or parasite
  • the activity of a compound of the invention against a microorganism may be measured by the minimum inhibitory concentration (MIC) of the compound in inhibiting the viability, growth, or replication of the microorganism.
  • the MIC of a compound of the invention is an MIC in inhibiting the viability the microorganism.
  • the MIC value of an inventive compound in inhibiting a microorganism is at most about 1 nM, at most about 3 nM, at most about 10 nM, at most about 30 nM, at most about 100 nM, at most about 300 nM, at most about 1 ⁇ M, at most about 3 ⁇ M, at most about 10 ⁇ M, at most about 30 ⁇ M, or at most about 100 ⁇ M.
  • the MIC value of an inventive compound in inhibiting a microorganism is at least about 1 nM, at least about 3 nM, at least about 10 nM, at least about 30 nM, at least about 100 nM, at least about 300 nM, at least about 1 ⁇ M, at least about 3 ⁇ M, at least about 10 ⁇ M, or at least about 30 ⁇ M.
  • MIC values are measured according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI) (which is incorporated herein by reference) (e.g., a broth microdilution method).
  • CCSI Clinical and Laboratory Standards Institute
  • MIC values are measured by a method described herein.
  • the activity of a compound of the invention against a microorganism may also be measured by the half maximal inhibitory concentration (IC 50 ) of the compound in inhibiting the viability, growth, or replication of the microorganism.
  • IC 50 half maximal inhibitory concentration
  • the IC 50 of a compound of the invention is an MIC in inhibiting the viability the microorganism.
  • the IC 50 value of an inventive compound in inhibiting a microorganism is at most about 1 nM, at most about 3 nM, at most about 10 nM, at most about 30 nM, at most about 100 nM, at most about 300 nM, at most about 1 ⁇ M, at most about 3 ⁇ M, at most about 10 ⁇ M, at most about 30 ⁇ M, or at most about 100 ⁇ M.
  • the IC 50 value of an inventive compound in inhibiting a microorganism is at least about 1 nM, at least about 3 nM, at least about 10 nM, at least about 30 nM, at least about 100 nM, at least about 300 nM, at least about 1 ⁇ M, at least about 3 ⁇ M, at least about 10 ⁇ M, or at least about 30 ⁇ M.
  • IC 50 values are measured according to the guidelines of the CLSI (e.g., a microdilution method). In certain embodiments, IC 50 values are measured by a method described herein.
  • the compounds of the invention may selectively inhibit the growth and/or reproduction of or kill a microorganism.
  • a compound of the invention is more active in inhibiting the growth and/or reproduction of or killing a first microorganism (e.g., a microorganism described herein) than in inhibiting the growth and/or reproduction of or killing a host cell.
  • a compound of the invention is more active in inhibiting the growth and/or reproduction of or killing a first microorganism than in inhibiting the growth and/or reproduction of or killing a second microorganism.
  • the selectivity of an inventive compound in inhibiting the growth and/or reproduction of or killing a first microorganism over a host cell or a second microorganism may be determined by the quotient of the MIC value of the inventive compound in inhibiting the growth and/or reproduction of or killing the host cell or second microorganism over the MIC value of the inventive compound in inhibiting the growth and/or reproduction of or killing the first microorganism.
  • the selectivity of an inventive compound in inhibiting the growth and/or reproduction of or killing a first microorganism over a host cell or a second microorganism may also be determined by the quotient of the IC 50 value of the inventive compound in inhibiting the growth and/or reproduction of or killing the host cell or second microorganism over the IC 50 value of the inventive compound in inhibiting the growth and/or reproduction of or killing the first microorganism.
  • the selectivity of an inventive compound in inhibiting the growth and/or reproduction of or killing a first microorganism over a host cell or a second microorganism is at least about 3-fold, at least about 10-fold, at least about 30-fold, at least about 100-fold, at least about 1,000-fold, at least about 10,000-fold, or at least about 100,000-fold.
  • the compounds of the invention may show low cytotoxicity toward mammalian cells (e.g., cytotoxicity IC 50 against HeLa cells being greater than 100 ⁇ M).
  • the compounds of the invention may show low hemolysis activity (e.g., not more than 1%, not more than 2%, not more than 4%, or not more than 6% hemolysis of red blood cells (RBCs) when treated with the compound at 200 ⁇ M).
  • compositions comprising a compound of the invention (e.g., a compound of Formula (I′) (e.g., Formula (I)), (II), or (III), or pharmaceutically acceptable salts thereof), and optionally an excipient (e.g., pharmaceutically acceptable excipient).
  • a compound of the invention e.g., a compound of Formula (I′) (e.g., Formula (I)), (II), or (III), or pharmaceutically acceptable salts thereof
  • an excipient e.g., pharmaceutically acceptable excipient
  • a composition of the invention is useful for disinfecting a surface.
  • the compound of the invention is provided in an effective amount in the composition.
  • the amount of the compound included in the composition is effective for killing at least 80%, at least 90%, at least 95%, at least 99%, at least 99.9%, or at least 99.99% of the microorganisms on the surface.
  • the amount of the compound included in the composition is effective for killing at most 90%, at most 95%, at most 99%, at most 99.9%, at most 99.99%, or at most 99.999% of the microorganisms on the surface.
  • a composition of the invention may include one or more excipients (e.g., water, detergent, bleach, surfactant) (e.g., pharmaceutically acceptable excipients).
  • a composition of the invention is a pharmaceutical composition comprising a compound of the invention and optionally a pharmaceutically acceptable excipient.
  • the compound of the invention is provided in an effective amount in the pharmaceutical composition.
  • the effective amount of the compound is a therapeutically effective amount.
  • the effective amount of the compound is a prophylactically effective amount.
  • the pharmaceutical compositions of the invention may be useful in the inventive methods.
  • the pharmaceutical compositions are useful in treating a microbial infection (e.g., a bacterial infection or mycobacterial infection).
  • the pharmaceutical compositions are useful in preventing a microbial infection (e.g., a bacterial infection or mycobacterial infection).
  • the pharmaceutical compositions are useful in inhibiting the growth of a microorganism (e.g., a microorganism described herein). In certain embodiments, the pharmaceutical compositions are useful in inhibiting the reproduction of a microorganism. In certain embodiments, the pharmaceutical compositions are useful in killing a microorganism. In certain embodiments, the pharmaceutical compositions are useful in inhibiting the formation and/or growth of a biofilm. In certain embodiments, the pharmaceutical compositions are useful in reducing or removing a biofilm. In certain embodiments, the pharmaceutical compositions are useful in disinfecting a surface. In certain embodiments, the pharmaceutical compositions are useful in cleaning a surface.
  • a microorganism e.g., a microorganism described herein.
  • the pharmaceutical compositions are useful in inhibiting the reproduction of a microorganism. In certain embodiments, the pharmaceutical compositions are useful in killing a microorganism. In certain embodiments, the pharmaceutical compositions are useful in inhibiting the formation and/or growth of a biofilm. In certain
  • compositions described herein can be prepared by any method known in the art of pharmacology.
  • preparatory methods include the steps of bringing the compound of the invention (the “active ingredient”) into association with a carrier and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping and/or packaging the product into a desired single- or multi-dose unit.
  • compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses.
  • a “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient.
  • the amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage, such as, for example, one-half or one-third of such a dosage.
  • Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered.
  • the composition may comprise between 0.1% and 100% (w/w) active ingredient.
  • compositions used in the manufacture of provided pharmaceutical compositions include inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may also be present in the composition.
  • Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.
  • Exemplary granulating and/or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood products, natural sponge, cation-exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.
  • crospovidone cross-linked poly(vinyl-pyrrolidone)
  • sodium carboxymethyl starch sodium starch glycolate
  • Exemplary surface active agents and/or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxy polymethylene, polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymer), carrageenan, cellulos
  • Exemplary binding agents include starch (e.g., cornstarch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (Veegum), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and/or mixtures
  • Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and other preservatives.
  • the preservative is an antioxidant.
  • the preservative is a chelating agent.
  • antioxidants include alpha tocopherol, ascorbic acid, acorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.
  • Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, and the like), citric acid and salts and hydrates thereof (e.g., citric acid monohydrate), fumaric acid and salts and hydrates thereof, malic acid and salts and hydrates thereof, phosphoric acid and salts and hydrates thereof, and tartaric acid and salts and hydrates thereof.
  • EDTA ethylenediaminetetraacetic acid
  • salts and hydrates thereof e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, and the like
  • citric acid and salts and hydrates thereof e.g., citric acid mono
  • antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.
  • antifungal preservatives include butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.
  • Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol.
  • Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.
  • preservatives include tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA), butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus, Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, and Euxyl.
  • Exemplary buffering agents include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer
  • Exemplary lubricating agents include magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.
  • Exemplary natural oils include almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, camomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton seed, emu, eucalyptus , evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba , macademia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasquana, savoury, sea
  • Exemplary synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixtures thereof.
  • Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may comprise inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • inert diluents commonly used in the art such as, for example, water or other solvents, so
  • the oral compositions can include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • the conjugates of the invention are mixed with solubilizing agents such as CremophorTM, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.
  • sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation can be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol.
  • acceptable vehicles and solvents that can be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid are used in the preparation of injectables.
  • the injectable formulations can be sterilized, for example, by filtration through a microbial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
  • compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing the conjugates of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient.
  • suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol mono
  • Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes.
  • Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
  • the active ingredient can be in a micro-encapsulated form with one or more excipients as noted above.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art.
  • the active ingredient can be admixed with at least one inert diluent such as sucrose, lactose or starch.
  • Such dosage forms may comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose.
  • the dosage forms may comprise buffering agents. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner.
  • opacifying agents include polymeric substances and waxes.
  • Dosage forms for topical and/or transdermal administration of a compound of this invention may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants and/or patches.
  • the active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier and/or any needed preservatives and/or buffers as can be required.
  • the present invention contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of an active ingredient to the body.
  • Such dosage forms can be prepared, for example, by dissolving and/or dispensing the active ingredient in the proper medium.
  • the rate can be controlled by either providing a rate controlling membrane and/or by dispersing the active ingredient in a polymer matrix and/or gel.
  • Suitable devices for use in delivering intradermal pharmaceutical compositions described herein include short needle devices.
  • Intradermal compositions can be administered by devices which limit the effective penetration length of a needle into the skin.
  • Jet injection devices which deliver liquid vaccines to the dermis via a liquid jet injector and/or via a needle which pierces the stratum corneum and produces a jet which reaches the dermis are suitable.
  • Ballistic powder/particle delivery devices which use compressed gas to accelerate the compound in powder form through the outer layers of the skin to the dermis are suitable.
  • conventional syringes can be used in the classical mantoux method of intradermal administration.
  • Formulations suitable for topical administration include, but are not limited to, liquid and/or semi liquid preparations such as liniments, lotions, oil-in-water and/or water-in-oil emulsions such as creams, ointments, and/or pastes, and/or solutions and/or suspensions.
  • Topically-administrable formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of the active ingredient can be as high as the solubility limit of the active ingredient in the solvent.
  • Formulations for topical administration may further comprise one or more of the additional ingredients described herein.
  • a pharmaceutical composition of the invention can be prepared, packaged, and/or sold in a formulation suitable for pulmonary administration via the buccal cavity.
  • a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 nanometers or from about 1 to about 6 nanometers.
  • Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant can be directed to disperse the powder and/or using a self-propelling solvent/powder dispensing container such as a device comprising the active ingredient dissolved and/or suspended in a low-boiling propellant in a sealed container.
  • Such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than 0.5 nanometers and at least 95% of the particles by number have a diameter less than 7 nanometers. Alternatively, at least 95% of the particles by weight have a diameter greater than 1 nanometer and at least 90% of the particles by number have a diameter less than 6 nanometers.
  • Dry powder compositions may include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form.
  • Low boiling propellants generally include liquid propellants having a boiling point of below 65° F. at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w/w) of the composition, and the active ingredient may constitute 0.1 to 20% (w/w) of the composition.
  • the propellant may further comprise additional ingredients such as a liquid non-ionic and/or solid anionic surfactant and/or a solid diluent (which may have a particle size of the same order as particles comprising the active ingredient).
  • compositions of the invention formulated for pulmonary delivery may provide the active ingredient in the form of droplets of a solution and/or suspension.
  • Such formulations can be prepared, packaged, and/or sold as aqueous and/or dilute alcoholic solutions and/or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization and/or atomization device.
  • Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, and/or a preservative such as methylhydroxybenzoate.
  • the droplets provided by this route of administration may have an average diameter in the range from about 0.1 to about 200 nanometers.
  • Formulations described herein as being useful for pulmonary delivery are useful for intranasal delivery of a pharmaceutical composition of the invention.
  • Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle from about 0.2 to 500 micrometers. Such a formulation is administered by rapid inhalation through the nasal passage from a container of the powder held close to the nares.
  • Formulations for nasal administration may, for example, comprise from about as little as 0.1% (w/w) to as much as 100% (w/w) of the active ingredient, and may comprise one or more of the additional ingredients described herein.
  • a pharmaceutical composition of the invention can be prepared, packaged, and/or sold in a formulation for buccal administration.
  • Such formulations may, for example, be in the form of tablets, and/or lozenges made using conventional methods, and may contain, for example, 0.1 to 20% (w/w) active ingredient, the balance comprising an orally dissolvable and/or degradable composition and, optionally, one or more of the additional ingredients described herein.
  • formulations for buccal administration may comprise a powder and/or an aerosolized and/or atomized solution and/or suspension comprising the active ingredient.
  • Such powdered, aerosolized, and/or aerosolized formulations when dispersed, may have an average particle and/or droplet size in the range from about 0.1 to about 200 nanometers, and may further comprise one or more of the additional ingredients described herein.
  • a pharmaceutical composition of the invention can be prepared, packaged, and/or sold in a formulation for ophthalmic administration.
  • Such formulations may, for example, be in the form of eye drops including, for example, a 0.1-1.0% (w/w) solution and/or suspension of the active ingredient in an aqueous or oily liquid carrier.
  • Such drops may further comprise buffering agents, salts, and/or one or more other of the additional ingredients described herein.
  • Other opthalmically-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form and/or in a liposomal preparation. Ear drops and/or eye drops are contemplated as being within the scope of this invention.
  • compositions suitable for administration to humans are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation.
  • compositions of the present invention are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions of the present invention will be decided by a physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts.
  • the compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), mucosal, nasal, bucal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol.
  • enteral e.g., oral
  • parenteral intravenous, intramuscular, intra-arterial, intramedullary
  • intrathecal subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal
  • topical as by powders, ointments, creams, and/or drops
  • mucosal nasal,
  • Specifically contemplated routes are oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration via blood and/or lymph supply, and/or direct administration to an affected site.
  • intravenous administration e.g., systemic intravenous injection
  • regional administration via blood and/or lymph supply
  • direct administration to an affected site.
  • the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract), and/or the condition of the subject (e.g., whether the subject is able to tolerate oral administration).
  • any two doses of the multiple doses include different or substantially the same amounts of a compound described herein.
  • the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses a day, two doses a day, one dose a day, one dose every other day, one dose every third day, one dose every week, one dose every two weeks, one dose every three weeks, or one dose every four weeks.
  • the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is one dose per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is two doses per day.
  • the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses per day.
  • the duration between the first dose and last dose of the multiple doses is one day, two days, four days, one week, two weeks, three weeks, one month, two months, three months, four months, six months, nine months, one year, two years, three years, four years, five years, seven years, ten years, fifteen years, twenty years, or the lifetime of the subject, tissue, or cell.
  • the duration between the first dose and last dose of the multiple doses is three months, six months, or one year.
  • the duration between the first dose and last dose of the multiple doses is the lifetime of the subject, tissue, or cell.
  • a dose (e.g., a single dose, or any dose of multiple doses) described herein includes independently between 0.1 ⁇ g and 1 ⁇ g, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg, between 0.1 mg and 1 mg, between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g, inclusive, of a compound described herein.
  • a dose described herein includes independently between 1 mg and 3 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 3 mg and 10 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 10 mg and 30 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 30 mg and 100 mg, inclusive, of a compound described herein.
  • dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult.
  • the amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult.
  • a dose described herein is a dose to an adult human whose body weight is 70 kg.
  • a compound or composition, as described herein can be administered in combination with one or more additional pharmaceutical agents.
  • the additional pharmaceutical agent is different from a compound of the invention, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.
  • the compounds or compositions can be administered in combination with additional pharmaceutical agents to improve their potency, efficacy, and/or bioavailability, reduce and/or modify their metabolism, inhibit their excretion, and/or modify their distribution within the body.
  • the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects.
  • the combination of a compound of the invention and an additional pharmaceutical agent shows a synergistic effect.
  • the compound or composition can be administered concurrently with, prior to, or subsequent to, one or more additional pharmaceutical agents, which are different from the compound or composition and may be useful as, e.g., combination therapies.
  • Pharmaceutical agents include therapeutically active agents.
  • Pharmaceutical agents also include prophylactically active agents.
  • Each additional pharmaceutical agent may be administered at a dose and/or on a time schedule determined for that pharmaceutical agent.
  • the additional pharmaceutical agents may also be administered together with each other and/or with the compound or composition described herein in a single dose or administered separately in different doses.
  • the particular combination to employ in a regimen will take into account compatibility of the inventive compound with the additional pharmaceutical agents and/or the desired therapeutic and/or prophylactic effect to be achieved. In general, it is expected that the additional pharmaceutical agents in combination be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually.
  • Exemplary additional pharmaceutical agents include, but are not limited to, antibiotics (e.g., antibacterial agents, antiviral agents, anti-fungal agents), anti-inflammatory agents, anti-pyretic agents, and pain-relieving agents.
  • Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved by the U.S.
  • the additional pharmaceutical agent is a chelator of a metal ion or metal atom.
  • the additional pharmaceutical agent is a chelator of a divalent metal ion (e.g., Mg(II), Ca(II), Sr(II), Mn(II), Fe(II), Co(II), Ni(II), Cu(II), or Zn(II)).
  • the additional pharmaceutical agent is a chelator of Cu(II), Mg(II), or Fe(II).
  • the additional pharmaceutical agent is di sodium 4, 5-dihydroxy-1,3-benzenedisulfonate (TIRON).
  • the additional pharmaceutical agent is 2,2′-dipyridyl, desferrioxamine (DFO, DESFERAL), deferasirox (EXJADE), deferiprone (L1, FERRIPROX), FERALEX-G, CaNa 3 DTPA, dexrazoxane, a phosphorothioate-oligonucleotide, desferrithiocin, or desazadesferrithiocin, or a derivative thereof.
  • the additional pharmaceutical agent is an antibiotic.
  • the additional pharmaceutical agent is an antibiotic effective against a microorganism described herein.
  • the additional pharmaceutical agent is an antibiotic effective against a bacterium.
  • the additional pharmaceutical agent is an antibiotic effective against a Gram-positive bacterium (e.g., a Staphylococcus species or Enterococcus species). In certain embodiments, the additional pharmaceutical agent is an antibiotic effective against a Gram-negative bacterium (e.g., an Acinetobacter species). In certain embodiments, the additional pharmaceutical agent is an antibiotic effective against a multidrug-resistant bacterium. In certain embodiments, the additional pharmaceutical agent is a ⁇ -lactam antibiotic.
  • the additional pharmaceutical agent is a penicillin (e.g., a penam, such as an aminopenicillin (e.g., amoxicillin, an ampicillin (e.g., pivampicillin, hetacillin, bacampicillin, metampicillin, talampicillin), epicillin), a carboxypenicillin (e.g., a carbenicillin (e.g., carindacillin), ticarcillin, temocillin), a ureidopenicillin (e.g., azlocillin, piperacillin, mezlocillin), a mecillinam (e.g., pivmecillinam), sulbenicillin, benzylpenicillin, clometocillin, benzathine benzylpenicillin, procaine benzylpenicillin, azidocillin, penamecillin, phenoxymethylpenicillin, propicillin, benzathine phenoxymethylpenicillin, phene
  • the additional pharmaceutical agent is an antiviral agent.
  • the additional pharmaceutical agent is ( ⁇ )-Oseltamivir, ⁇ -D-ribofuranose, 1-acetate 2,3,5-tribenzoate, 1-Docosanol, 2-Amino-6-chloropurine, 5-Iodo-2′-deoxyuridine, 6-Chloropurine, Abacavir sulfate, Abacavir-epivir mixt., Acyclovir, Acyclovir sodium, Adefovir dipivoxil, Amantadine (e.g., Amantadine hydrochloride), Amantadine hydrochloride, anti-HIV agent (e.g., Abacavir, Amprenavir, Atazanavir, Azidothymidine, Bryostatin (e.g., Bryostatin 1, Bryostatin 10, Bryostatin 11, Bryostatin 12, Bryostatin 13, B
  • the additional pharmaceutical agent is a fungicide.
  • the additional pharmaceutical agent is ( ⁇ )-Fumagillin, ( ⁇ )-Metalaxyl, 1,2, 5-Fluorocytosine, Acrisorcin, Anilazine, Antifouling agent, Azoxystrobin, Benomyl, Bordeaux mixture, Captan, Carbendazim, Caspofungin acetate, Chlorothalonil, Clotrimazole, Dichlofluanid, Dinocap, Dodine, Fenhexamid, Fenpropimorph, Ferbam, Fluconazole, Fosetyl Al, Griseofulvin, Guanidine (e.g., Agmatine, Amiloride hydrochloride, Biguanide (e.g., Imidodicarbonimidic diamide, N,N-dimethyl-,hydrochloride (1:1) (e.g., Metformin hydrochloride), Metformin), Cimetidine, Guane
  • the additional pharmaceutical agent is a protozoacide.
  • the additional pharmaceutical agent is Amebicide, Antimalarial (e.g., Artemisinin, Chloroquine (e.g., Chloroquine phosphate), Mefloquine, Sulfadoxine), Coccidiostat, Leishmanicide, Trichomonacide, or Trypanosomicide (e.g., Eflornithine).
  • the additional pharmaceutical agent is a parasiticide.
  • the additional pharmaceutical agent is antihelmintic (e.g., Abamectin, Dimethylformocarb othialdine, Niclosamide, Schistosomicide), protozoacide (e.g., Amebicide, antimalarial (e.g., Artemisinin, chloroquine (e.g., chloroquine phosphate), Mefloquine, Sulfadoxine), coccidiostat, leishmanicide, trichomonacide, or trypanosomicide (e.g., Eflornithine)).
  • antihelmintic e.g., Abamectin, Dimethylformocarb othialdine, Niclosamide, Schistosomicide
  • protozoacide e.g., Amebicide
  • antimalarial e.g., Artemisinin
  • chloroquine e.g., chloroquine phosphate
  • Mefloquine e.g.,
  • the pharmaceutical composition is substantially free (e.g., at least 70% free, at least 80% free, at least 90% free, at least 95% free, at least 99% free, or at least 99.9% free) of a metal ion or metal atom.
  • the pharmaceutical composition is substantially free of a divalent metal ion (e.g., Mg(II), Ca(II), Sr(II), Mn(II), Fe(II), Co(II), Ni(II), Cu(II), or Zn(II)).
  • the pharmaceutical composition is substantially free of Cu(II), Mg(II), or Fe(II).
  • kits e.g., pharmaceutical packs.
  • the kits provided may comprise a compound or composition (e.g., pharmaceutical composition) of the invention and a container (e.g., a vial, ampule, bottle, syringe, dispenser package, tube, inhaler, and/or other suitable container).
  • a kit of the invention further includes a second container comprising an excipient (e.g., pharmaceutically acceptable excipient) for dilution or suspension of an inventive compound or composition.
  • the compound or composition of the invention provided in a first container and a second container are combined to form one unit dosage form.
  • kits including a first container comprising a compound or composition of the invention.
  • a kit of the invention includes a first container comprising a compound of Formula (I′) (e.g., Formula (I)), (II), or (III), or a pharmaceutically acceptable salt thereof, or a composition thereof.
  • kits are useful in treating a microbial infection in a subject in need thereof. In certain embodiments, the kits are useful in preventing a microbial infection in a subject in need thereof. In certain embodiments, the microbial infection is a bacterial infection. In certain embodiments, the bacterial infection is an infection caused by a Gram-positive bacterium. In certain embodiments, the bacterial infection is an infection caused by a Gram-negative bacterium. In certain embodiments, the microbial infection is a mycobacterial infection. In certain embodiments, the kits are useful in inhibiting the growth of a microorganism. In certain embodiments, the kits are useful in inhibiting the reproduction of a microorganism.
  • kits are useful in killing a microorganism. In certain embodiments, the kits are useful in inhibiting the formation and/or growth of a biofilm. In certain embodiments, the kits are useful in reducing or removing a biofilm. In certain embodiments, the kits are useful in disinfecting a surface. In certain embodiments, the kits are useful for screening a library of compounds to identify a compound that is useful in the methods of the invention.
  • the kit further includes instructions for using the compound or pharmaceutical composition included in the kit (e.g., for administering to a subject in need of treatment of a microbial infection a compound or pharmaceutical composition of the invention, for contacting a microorganism with a compound or pharmaceutical composition of the invention, or for contacting a biofilm with a compound or pharmaceutical composition of the invention).
  • the kits may also include information as required by a regulatory agency such as the U.S. Food and Drug Administration (FDA).
  • the information included in the kits is prescribing information.
  • the kits and instructions provide for treating a microbial infection in a subject in need thereof.
  • the kits and instructions provide for preventing a microbial infection in a subject in need thereof.
  • kits and instructions provide for inhibiting the growth of a microorganism. In certain embodiments, the kits and instructions provide for inhibiting the reproduction of a microorganism. In certain embodiments, the kits and instructions provide for killing a microorganism. In certain embodiments, the kits and instructions provide for inhibiting the formation and/or growth of a biofilm. In certain embodiments, the kits and instructions provide for reducing or removing a biofilm. In certain embodiments, the kits and instructions provide for disinfecting a surface. In certain embodiments, the kits and instructions provide for screening a library of compounds to identify a compound that is useful in the methods of the invention.
  • the kit of the invention may include one or more additional agents described herein (e.g., additional pharmaceutical agents) as a separate composition.
  • non-replicative bacteria e.g., dormant persister cells, bacterial biofilms
  • bacterial biofilms surface-attached bacterial communities with altered physiologies, gene expression profiles and growth-rates
  • persister cells 6 has grown considerably, yet the ability to target persistent bacterial phenotypes remains an unmet challenge.
  • innovative strategies to identify antibacterial agents that operate through growth-independent mechanisms may be employed.
  • Biofilm-eradicating agents typically operate through the disruption of bacterial membranes (e.g., antimicrobial peptides, 8,9 quaternary ammonium cations/QACs 10 ). Although these compounds are indeed valuable, new biofilm-eradicating agents with complementary modes of action are of great importance and have multiple therapeutic applications to address persistent bacterial infections.
  • HP 202 a synthetic analogue of marine phenazine antibiotic 201
  • MBEC biofilm eradication concentration
  • the mechanism of 202 was investigated, since 2-bromo-1-hydroxyphenazine 201 belongs to the family of redox-active phenazine antibiotics produced by Pseudomonas and Streptomyces bacteria. 18 In addition, one of the goals was to synthesize more efficient biofilm-eradicating agents, target persister cells in non-biofilm cultures and evaluate HP analogues against the slow-growing pathogen Mycobacterium tuberculosis (MtB). Compounds that can effectively eradicate biofilms, persister cells and MtB are promising agents to address problems associated with bacterial persistence.
  • MtB Mycobacterium tuberculosis
  • the present invention also provides methods for treating a microbial infection (e.g., bacterial infection or mycobacterial infection) in a subject in need thereof.
  • a microbial infection e.g., bacterial infection or mycobacterial infection
  • the microbial infection is treated by the inventive methods.
  • the present invention further provides methods for preventing a microbial infection (e.g., bacterial infection or mycobacterial infection) in a subject in need thereof.
  • the microbial infection is prevented by the inventive methods.
  • the subject described herein is an animal. In certain embodiments, the subject is a non-human animal. In certain embodiments, the subject is a mammal. In certain embodiments, the subject is a human. In certain embodiments, the subject is a human with cystic fibrosis. In certain embodiments, the subject is a non-human mammal. In certain embodiments, the subject is a domesticated animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a companion animal, such as a dog or cat. In certain embodiments, the subject is a livestock animal, such as a cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a zoo animal. In another embodiment, the subject is a research animal, such as a rodent, dog, or non-human primate. In certain embodiments, the subject is a non-human transgenic animal, such as a transgenic mouse or transgenic pig.
  • the methods of the invention include administering to a subject in need thereof an effective amount of a compound or pharmaceutical composition of the invention.
  • the methods of the invention include administering to a subject in need thereof an effective amount of a compound of Formula (I′) (e.g., Formula (I)), (II), or (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the methods of the invention include administering to a subject in need thereof a therapeutically effective amount of a compound of the invention, or a pharmaceutical composition thereof.
  • the methods of the invention include administering to a subject in need thereof a prophylactically effective amount of a compound of the invention, or a pharmaceutical composition thereof.
  • the microbial infection that is treated and/or prevented by the inventive methods or using the inventive compounds or pharmaceutical compositions thereof is caused by a multidrug-resistant microorganism and/or a microorganism resistant to methicillin, penicillin, ciprofloxacin, rifampin, vancomycin, daptomycin, linezolid, an antibiotic described herein, or a combination thereof.
  • the microbial infection is a microbial respiratory tract infection.
  • the microbial infection is microbial pneumonia.
  • the microbial infection is microbial sinusitis.
  • the microbial infection is tuberculosis (TB).
  • the microbial infection is microbial Crohn's disease, paratuberculosis, Buruli ulcer, leprosy, or aquarium granuloma.
  • the microbial infection is a microbial gastrointestinal tract infection.
  • the microbial infection is microbial diarrhea.
  • the microbial infection is a microbial urogenital tract infection.
  • the microbial infection is a microbial bloodstream infection.
  • the microbial infection is microbial hemolytic uremic syndrome.
  • the microbial infection is microbial endocarditis.
  • the microbial infection is a microbial ear infection.
  • the microbial infection is a microbial skin infection (e.g., microbial acne vulgaris). In certain embodiments, the microbial infection is a microbial oral infection. In certain embodiments, the microbial infection is a microbial dental infection. In certain embodiments, the microbial infection is gingivitis. In certain embodiments, the microbial infection is dental plaque caused by a microorganism. In certain embodiments, the microbial infection is microbial meningitis. In certain embodiments, the microbial infection is a microbial wound or surgical site infection. In certain embodiments, the microbial infection is a microbial burn wound infection.
  • the microbial infection is a microbial infection associated with cystic fibrosis. In certain embodiments, the microbial infection is a microbial infection associated with an implanted device. In certain embodiments, the microbial infection is a microbial infection associated with a dental implant. In certain embodiments, the microbial infection is a microbial infection associated with a catheter. In certain embodiments, the microbial infection is a microbial infection associated with a heart valve. In certain embodiments, the microbial infection is a microbial infection associated with an intrauterine device. In certain embodiments, the microbial infection is a microbial infection associated with a joint prosthesis. In certain embodiments, the microbial infection is a bacterial infection.
  • the bacterial infection is caused by a Gram-positive bacterium (e.g., a Gram-positive bacterium described herein). In certain embodiments, the bacterial infection is caused by a Gram-negative bacterium (e.g., a Gram-negative bacterium described herein). In certain embodiments, the bacterial infection is caused by a multidrug-resistant bacterium. In certain embodiments, the bacterial infection is caused by a strain of Staphylococcus aureus . In certain embodiments, the bacterial infection is a methicillin-resistant Staphylococcus aureus (MRSA)-related infection.
  • MRSA methicillin-resistant Staphylococcus aureus
  • the bacterial infection is caused by a strain of Staphylococcus epidermidis (e.g., MRSE). In certain embodiments, the bacterial infection is an MRSE-related infection. In certain embodiments, the bacterial infection is caused by a strain ofEnterococcusfaecium. In certain embodiments, the bacterial infection is caused by Acinetobacter baumannii ( A. baumannii ). In certain embodiments, the microbial infection is a mycobacterial infection. In certain embodiments, the microbial infection is caused by a mycobacterium (e.g., a strain of Mycobacterium tuberculosis ). In certain embodiments, the microbial infection is caused by an archaeon.
  • MRSE Staphylococcus epidermidis
  • the bacterial infection is an MRSE-related infection.
  • the bacterial infection is caused by a strain ofEnterococcusfaecium.
  • the bacterial infection is caused by Acinetobacter baumannii
  • the microbial infection is caused by a protist. In certain embodiments, the microbial infection is caused by a protozoon. In certain embodiments, the microbial infection is caused by an alga. In certain embodiments, the microbial infection is caused by a fungus. In certain embodiments, the microbial infection is caused by yeast. In certain embodiments, the microbial infection is caused by a mold. In certain embodiments, the microbial infection is caused by a parasite. In certain embodiments, the microbial infection is a microbial infection associated with a biofilm.
  • Another aspect of the present invention relates to methods of inhibiting the growth of a microorganism using a compound of the invention, or a pharmaceutical composition thereof.
  • an inventive method selectively inhibits the growth of a first microorganism (e.g., a microorganism described herein), compared to the inhibition of the growth of a host cell or a second microorganism.
  • the growth of a microorganism is inhibited by the inventive methods.
  • the growth of a first microorganism is selectively inhibited by the inventive methods, compared to the inhibition of the growth of a host cell or a second microorganism.
  • Another aspect of the present invention relates to methods of inhibiting the reproduction of a microorganism using a compound of the invention, or a pharmaceutical composition thereof.
  • an inventive method selectively inhibits the reproduction of a first microorganism (e.g., a microorganism described herein), compared to the inhibition of the reproduction of a host cell or a second microorganism.
  • the reproduction of a microorganism is inhibited by the inventive methods.
  • the reproduction of a first microorganism is selectively inhibited by the inventive methods, compared to the inhibition of the reproduction of a host cell or a second microorganism.
  • Another aspect of the present invention relates to methods of inhibiting the viability of a microorganism using a compound of the invention, or a pharmaceutical composition thereof.
  • an inventive method selectively inhibits the viability of a first microorganism (e.g., a microorganism described herein), compared to the inhibition of the viability of a host cell or a second microorganism.
  • the viability of a microorganism is inhibited by the inventive methods.
  • the viability of a first microorganism is selectively inhibited by the inventive methods, compared to the inhibition of the viability of a host cell or a second microorganism.
  • Another aspect of the present invention relates to methods of killing a microorganism using a compound of the invention, or a pharmaceutical composition thereof.
  • an inventive method selectively kills a first microorganism (e.g., a microorganism described herein), compared to the killing of a host cell or a second microorganism.
  • a microorganism is killed by the inventive methods.
  • a first microorganism is selectively killed by the inventive methods, compared to the killing of a host cell or a second microorganism.
  • the methods of the invention include contacting a microorganism with an effective amount of a compound or pharmaceutical composition of the invention.
  • the methods of the invention include contacting a microorganism with an effective amount of a compound of Formula (I′) (e.g., Formula (I)), (II), or (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the methods of the invention include contacting a microorganism with a therapeutically effective amount of a compound of the invention, or a pharmaceutical composition thereof.
  • the methods of the invention include contacting a microorganism with a prophylactically effective amount of a compound of the invention, or a pharmaceutical composition thereof.
  • a biofilm is typically formed on a living or non-living, solid or liquid surface.
  • a biofilm is formed on the surface of a biological sample (e.g., a tooth, oral soft tissue, middle ear, gastrointestinal tract, urogenital tract, respiratory tract, or eye).
  • a biofilm is formed on the surface of an implanted device (e.g., a dental implant, catheter, heart valve, intrauterine device, or joint prosthesis).
  • the biofilm is in vitro.
  • the biofilm is in vivo.
  • the biofilm described herein comprises a microorganism.
  • the biofilm comprises a microorganism (e.g., bacterium).
  • the biofilm comprises a strain of Staphylococcus aureus (e.g., a methicillin-resistant strain of Staphylococcus aureus ).
  • the biofilm comprises a strain of Staphylococcus epidermidis (e.g., a strain of MRSE). Free-floating microorganisms may accumulate on a surface, and the resulting biofilm may grow.
  • the concentration of microorganisms may be high and/or the resistance of the microorganisms in the biofilm to antimicrobial agents may be high.
  • Antimicrobials may be inactivated or fail to penetrate into the biofilm. Therefore, microbial infections associated with a biofilm (e.g., microbial infections caused by a biofilm) are typically more difficult to treat than microbial infections not associated with a biofilm.
  • Another aspect of the present invention relates to methods of inhibiting the formation of a biofilm using a compound of the invention, or a pharmaceutical composition thereof.
  • the formation of a biofilm is inhibited by the inventive methods.
  • Another aspect of the present invention relates to methods of inhibiting the growth of a biofilm using a compound of the invention, or a pharmaceutical composition thereof.
  • the growth of a biofilm is inhibited by the inventive methods.
  • a biofilm is reduced by the inventive methods, e.g., reduced by at least 10%, at least 20%, at least 30%, at least 50%, at least 70%, at least 90%, at least 99%, at least 99.9%, or at least 99.99%, in terms of the volume of the biofilm.
  • a biofilm is reduced by the inventive methods by not more than 10%, not more than 20%, not more than 30%, not more than 50%, not more than 70%, not more than 90%, not more than 99%, not more than 99.9%, or not more than 99.99%, in terms of the volume of the biofilm.
  • a biofilm is reduced by the inventive methods by at least 10%, at least 20%, at least 30%, at least 50%, at least 70%, at least 90%, at least 99%, at least 99.9%, or at least 99.99%, in terms of the number of microorganisms (e.g., bacteria) in the biofilm.
  • a biofilm is reduced by the inventive methods by not more than 10%, not more than 20%, not more than 30%, not more than 50%, not more than 70%, not more than 90%, not more than 99%, not more than 99.9%, or not more than 99.99%, in terms of the number of microorganisms (e.g., bacteria) in the biofilm.
  • Another aspect of the present invention relates to methods of removing a biofilm (e.g., eradicating a biofilm (e.g., reducing the volume of the biofilm by at least 99% and/or killing essentially all (e.g., at least 99%) of the microorganisms (e.g., bacteria) in the biofilm)) using a compound of the invention, or a pharmaceutical composition thereof.
  • a biofilm is removed by the inventive methods.
  • a biofilm reduced or removed by a method of the invention does not regrow one day, two days, four days, one week, two weeks, three weeks, or one month subsequent to the biofilm being subject to the method.
  • the methods of the invention include contacting a biofilm with an effective amount of a compound or pharmaceutical composition of the invention.
  • the methods of the invention include contacting a biofilm with an effective amount of a compound of Formula (I′) (e.g., Formula (I)), (II), or (III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.
  • the methods of the invention include contacting a biofilm with a therapeutically effective amount of a compound of the invention, or a pharmaceutical composition thereof.
  • the methods of the invention include contacting a biofilm with a prophylactically effective amount of a compound of the invention, or a pharmaceutical composition thereof.
  • Another aspect of the present invention relates to methods of disinfecting a surface, the methods including contacting the surface with an effective amount of a compound or composition (e.g., pharmaceutical composition) of the invention.
  • a compound or composition e.g., pharmaceutical composition
  • the number of viable microorganisms on the surface is reduced after the surface is contacted with the compound or composition.
  • the surface is a biological surface, such as skin (e.g., skin of: the hands, feet, arms, legs, face, neck, torso, or cavity (e.g., oral cavity)) of a subject.
  • the surface is a non-biological surface (e.g., a surface in a household, industrial, or medical setting, such as a surface of: a kitchen, bathroom, table top, floor, wall, window, utensil, cutlery, crockery, or medical device).
  • a non-biological surface may be a surface of a solid material, such as plastic, wood, bamboo, metal, ceramic, glass, concrete, stone, paper, fabric, or a combination thereof.
  • a non-biological surface may be painted or non-painted, or coated or non-coated.
  • the amount of the compound or composition is effective for killing at least 80%, at least 90%, at least 95%, at least 99%, at least 99.9%, or at least 99.99% of the microorganisms on the surface.
  • the microorganism described herein is a bacterium.
  • the microorganism is multidrug-resistant.
  • the microorganism is resistant to methicillin, penicillin, ciprofloxacin, rifampin, vancomycin, daptomycin, linezolid, or a combination thereof.
  • the microorganism is associated with a biofilm (e.g., present in and/or on a biofilm, able to form a biofilm, and/or able to increase the size of a biofilm).
  • the bacterium is a Gram-positive bacterium.
  • the bacterium is a multidrug-resistant bacterium.
  • the bacterium is a Staphylococcus species. In certain embodiments, the bacterium is a Staphylococcus aureus ( S. aureus ) strain (e.g., ATCC 25923). In certain embodiments, the bacterium is methicillin-resistant Staphylococcus aureus (MRSA). In certain embodiments, the bacterium is the methicillin-resistant Staphylococcus aureus clinical isolate (MRSA-2, a clinical isolate from a patient treated at Shands Hospital; obtained from the Emerging Pathogens Institute at the University of Florida), such as the methicillin-resistant Staphylococcus aureus clinical isolate reported in Abouelhassan et al., Bioorg. Med.
  • MRSA methicillin-resistant Staphylococcus aureus clinical isolate
  • the bacterium is a Staphylococcus epidermidis ( S. epidermidis ) strain (e.g., ATCC 12228 or ATCC 35984). In certain embodiments, the bacterium is an MRSE strain.
  • the bacterium is a Staphylococcus auricularis, Staphylococcus carnosus, Staphylococcus condimenti, Staphylococcus massiliensis, Staphylococcus piscifermentans, Staphylococcus simulans, Staphylococcus capitis, Staphylococcus caprae, Staphylococcus saccharolyticus, Staphylococcus devriesei, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus chromogenes, Staphylococcus felis, Staphylococcus delphini, Staphylococcus hyicus, Staphylococcus intermedius, Staphylococcus lutrae, Staphylococcus microti, Staphylococcus muscae, Staphylococcus pseudintermedius, Staphylococcus
  • the bacterium is a Streptococcus species. In certain embodiments, the bacterium is a Streptococcus agalactiae, Streptococcus anginosus, Streptococcus bovis, Streptococcus canis, Streptococcus constellatus, Streptococcus dysgalactiae, Streptococcus equinus, Streptococcus iniae, Streptococcus intermedius, Streptococcus mitis, Streptococcus mutans, Streptococcus oralis, Streptococcus parasanguinis, Streptococcus peroris, Streptococcus pneumoniae, Streptococcus pseudopneumoniae, Streptococcus pyogenes, Streptococcus ratti, Streptococcus salivarius, Streptococcus tigurinus, Streptococcus thermo
  • the bacterium is an Enterococcus species. In certain embodiments, the bacterium is an Enterococcus avium, Enterococcus durans , Enterococcusfaecalis, Enterococcus faecium, Enterococcus gallinarum, Enterococcus hirae , or Enterococcus solitarius strain. In certain embodiments, the bacterium is an Enterococcus faecium strain (e.g., a vancomycin-resistant strain of Enterococcus faecium (VRE); ATCC 700221). In certain embodiments, the bacterium is a Listeria species.
  • VRE vancomycin-resistant strain of Enterococcus faecium
  • the bacterium is a Listeria yakmannii, Listeria grayi, Listeria innocua, Listeria ivanovii, Listeria marthii, Listeria monocytogenes, Listeria rocourtiae, Listeria seeligeri, Listeria weihenstephanensis , or Listeria welshimeri strain. In certain embodiments, the bacterium is a Clostridium species.
  • the bacterium is a Clostridium acetobutylicum, Clostridium argentinense, Clostridium aerotolerans, Clostridium baratii, Clostridium beijerinckii, Clostridium bifermentans, Clostridium botulinum, Clostridium butyricum, Clostridium cadaveris, Clostridium cellulolyticum, Clostridium chauvoei, Clostridium clostridioforme, Clostridium colicanis, Clostridium difficile, Clostridium estertheticum , Clostridiumfallax, Clostridium feseri, Clostridium formicaceticum, Clostridium histolyticum, Clostridium innocuum, Clostridium kluyveri, Clostridium ljungdahlii, Clostridium lavalense, Clostridium leptum, Clostridium novyi, Clostridium
  • the bacterium is a Gram-negative bacterium. In certain embodiments, the bacterium is a bacterium described herein, provided that the bacterium is not a Gram-negative bacterium. In certain embodiments, the Gram-negative bacterium is an Escherichia species. In certain embodiments, the Gram-negative bacterium is an Escherichia coli ( E. coli ) strain (e.g., ATCC 33475, K-12, CFT073, ATCC 43895).
  • the Gram-negative bacterium is an Escherichia albertii strain, Escherichia blattae strain, Escherichia fergusonii strain, Escherichia hermannii strain, or Escherichia vulneris strain. In certain embodiments, the Gram-negative bacterium is a Pseudomonas species. In certain embodiments, the Gram-negative bacterium is a Pseudomonas aeruginosa strain.
  • the Gram-negative bacterium is a Pseudomonas alcaligenes strain, Pseudomonas anguilliseptica strain, Pseudomonas argentinensis strain, Pseudomonas borbori strain, Pseudomonas citronellolis strain, Pseudomonas flavescens strain, Pseudomonas mendocina strain, Pseudomonas nitroreducens strain, Pseudomonas oleovorans strain, Pseudomonas pseudoalcaligenes strain, Pseudomonas resinovorans strain, Pseudomonas straminea strain, Pseudomonas chlororaphis strain, Pseudomonas fluorescens strain, Pseudomonas pertucinogena strain, Pseudomonas putida strain
  • the Gram-negative bacterium is a Klebsiella species. In certain embodiments, the Gram-negative bacterium is a Klebsiella granulomatis strain, Klebsiella oxytoca strain, Klebsiella pneumoniae strain, Klebsiella terrigena strain, or Klebsiella planticola strain. In certain embodiments, the Gram-negative bacterium is a strain ofKlebsiella pneumoniae ( K. pneumoniae ). In certain embodiments, the Gram-negative bacterium is a Salmonella species. In certain embodiments, the Gram-negative bacterium is a Salmonella bongori strain or Salmonella enterica strain, e.g., Salmonella typhi .
  • the Gram-negative bacterium is an Acinetobacter species. In certain embodiments, the Gram-negative bacterium is an Acinetobacter baumannii strain. In certain embodiments, the Gram-negative bacterium is an Acinetobacter baylyi strain, Acinetobacter bouvetii strain, Acinetobacter calcoaceticus strain, Acinetobacter organizerri strain, Acinetobacter grimontii strain, Acinetobacter haemolyticus strain, Acinetobacter johnsonii strain, Acinetobacter junii strain, Acinetobacter lwoffii strain, Acinetobacter parvus strain, Acinetobacter pittii strain, Acinetobacter radioresistens strain, Acinetobacter schindleri strain, Acinetobacter tandoii strain, Acinetobacter tjernbergiae strain, Acinetobacter towneri strain, Acinetobacter ursingii strain, or Acinetobacter gyllenbergii strain.
  • the microorganism is a mycobacterium . In certain embodiments, the microorganism is a strain of Mycobacterium tuberculosis . In certain embodiments, the microorganism is a strain of: Mycobacterium bovis, Mycobacterium bovis BCG, Mycobacterium africanum, Mycobacterium canetti, Mycobacterium caprae, Mycobacterium microti, Mycobacterium Pinnipedii, Mycobacterium avium, Mycobacterium avium paratuberculosis, Mycobacterium avium silvaticum, Mycobacterium avium hominissuis, Mycobacterium colombiense, Mycobacterium indicus pranii, Mycobacterium gastri, Mycobacterium kansasii, Mycobacterium hiberniae, Mycobacterium nonchromogenicum, Mycobacterium terrae, Mycobacterium triviale, Mycobacterium ulcerans, Mycobacterium pseudoshot
  • the microorganism described herein is an archaeon. In certain embodiments, the microorganism is a protist. In certain embodiments, the microorganism is a protozoon. In certain embodiments, the microorganism is an alga. In certain embodiments, the microorganism is a fungus. In certain embodiments, the microorganism is yeast. In certain embodiments, the microorganism is a mold. In certain embodiments, the microorganism is a parasite.
  • the microorganism described herein is in vitro. In certain embodiments, the microorganism is in vivo.
  • a method of the invention is an in vitro method. In certain embodiments, a method of the invention is an in vivo method.
  • the present invention provides uses of the compounds, compositions, and pharmaceutical compositions of the invention for manufacturing a medicament for treating a microbial infection (e.g., bacterial infection or mycobacterial infection).
  • a microbial infection e.g., bacterial infection or mycobacterial infection.
  • the present invention provides uses of the compounds, compositions, and pharmaceutical compositions of the invention for manufacturing a medicament for preventing a microbial infection (e.g., bacterial infection or mycobacterial infection).
  • a microbial infection e.g., bacterial infection or mycobacterial infection.
  • the present invention provides the compounds, compositions, and pharmaceutical compositions of the invention for use in treating a microbial infection (e.g., bacterial infection or mycobacterial infection).
  • a microbial infection e.g., bacterial infection or mycobacterial infection.
  • the present invention provides the compounds, compositions, and pharmaceutical compositions of the invention for use in preventing a microbial infection (e.g., bacterial infection or mycobacterial infection).
  • a microbial infection e.g., bacterial infection or mycobacterial infection.
  • the compounds provided herein can be prepared from readily available starting materials using the following general methods and procedures. It will be appreciated that where typical or preferred process conditions (e.g., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by those skilled in the art by routine optimization procedures.
  • protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions.
  • suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are well known in the art. For example, numerous protecting groups, and their introduction and removal, are described in Greene et al., Protecting Groups in Organic Synthesis , Second Edition, Wiley, New York, 1991, and references cited therein.
  • the compounds of the invention can be prepared using previously reported synthetic protocols (e.g., E. Breitmaier, J Org. Chem., 1976, 41, 2104-2108; D. L. Vivan, Nature, 1956, 178, 753; M. Conda-Sheridan et al., J. Med. Chem., 2010, 53, 8688-8699; G. W. Rewcastle et al., J. Med. Chem., 1987, 30, 843-851; international PCT application publication, WO 2015/100331, published Jul. 2, 2015; each of which is incorporated herein by reference).
  • select compounds of the invention were prepared according to the methods shown in Schemes 1 and 2.
  • a library of five HP analogues was prepared that contained various substitutions in the 7- and 8-position of the phenazine related to previous work by Cushman 19 (Scheme 2).
  • Demethylation of 1-methoxyphenazines 203-205 proceeded smoothly using boron tribromide (BBr 3 ) to afford 1-hydroxyphenazines 206-208 (78-99%), which were then brominated using N-bromosuccinimide (NBS) to yield 209-211 (38-84% yield).
  • BBr 3 boron tribromide
  • NBS N-bromosuccinimide
  • 1-Methoxyphenazine 203 was selectively iodinated at the 4-position using sodium periodate (NaIO 4 )/potassium iodide (KI)/sodium chloride (NaCl) to afford 212 (59% yield), followed by demethylation using BBr 3 to give 213 (97% yield).
  • 3-methoxycatechol 215 (1.40 g, 10 mmol) was dissolved in diethyl ether (30 mL), then cooled to ⁇ 78° C. Tetrachloro-o-benzoquinone (3.07 g, 12.5 mmol) was then added, and the solution and allowed to stir at ⁇ 78° C. for 4 hours. The reaction mixture was then filtered twice under reduced pressure to afford benzoquinone 216 as a dark brown solid, which was used without further purification.
  • 7,8-Dichloro-4-iodo-1-hydroxyphenazine 213 (115 mg, 0.296 mmol) and N-bromosuccinimide (52.6 mg, 0.296 mmol) were dissolved in 20 mL of dichloromethane, and the resulting mixture was allowed to stir at room temperature for 1 hour. The reaction contents were then concentrated, adsorbed onto silica gel, and purified via column chromatography using dichloromethane to elute the product, which was obtained as an orange solid (32%, 43.6 mg).
  • Example 1 The references cited in Example 1 are included in Example 2. In Table 1, the different compound numbers in the same cell of Table 1 refer to the same compound.
  • MIC minimum inhibitory concentration for each compound described herein was determined by the broth microdilution method as recommended by the Clinical and Laboratory Standards Institute (CLSI) (Clinical and Laboratory Standards Institute. 2009. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard, eighth edition (M7-A8)).
  • CLSI Clinical and Laboratory Standards Institute
  • M7-A8 Clinical and Laboratory Standards Institute. 2009. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard, eighth edition (M7-A8)).
  • CLSI Clinical and Laboratory Standards Institute. 2009. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard, eighth edition (M7-A8)).
  • CLSI Clinical and Laboratory Standards Institute. 2009. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard, eighth edition (M7-A8)).
  • M7-A8 Clinical and Laboratory Standards Institute. 2009. Methods for dilution antimicrobial susceptibility tests
  • the MIC was defined as the lowest concentration of a compound that prevented bacterial growth after incubating of 16 to 18 hours at 37° C. (MIC values were supported by spectrophotometric readings at OD 600 ). The concentration range tested for each compound during this study was 0.10 to 100 ⁇ M.
  • DMSO served as the vehicle and negative control in each microdilution MIC assay. DMSO was serially diluted at the same concentration as the compounds with a top concentration of 1% v/v.
  • MRSA methicillin-resistant Staphylococcus aureus
  • MRSA-1 methicillin-resistant Staphylococcus epidermidis
  • MRSE strain ATCC 35984 methicillin-resistant Staphylococcus epidermidis
  • VRE vancomycin-resistant Enterococcus faecium
  • cytotoxicities of select compounds of the invention against HeLa cells were determined. HeLa cytotoxicity was assessed using the LDH release assay described by CYTOTOX96 (Promega G1780). HeLa cells were grown in Dulbecco's Modified Eagle Medium (DMEM; Gibco) supplemented with 10% Fetal Bovine Serum (FBS) at 37° C. with 5% CO 2 . When the HeLa cultures exhibited 70-80% confluence, halogenated phenazines were then diluted by DMEM (10% FBS) at concentrations of 25, 50, and 100 ⁇ M and added to HeLa cells.
  • DMEM Dulbecco's Modified Eagle Medium
  • FBS Fetal Bovine Serum
  • Triton X-100 (at 2% v/v) was used as the positive control for maximum lactate dehydrogenate (LDH) activity in this assay (e.g., complete cell death), while “medium only” lanes served as negative control lanes (e.g., no cell death).
  • LDH lactate dehydrogenate
  • DMSO was used as the vehicle control.
  • HeLa cells were treated with compounds for 24 hours, and then 50 ⁇ L of the supernatant was transferred into a fresh 96-well plate where 50 ⁇ L of the reaction mixture was added to the 96-well plate and incubated at room temperature for 30 minutes. Finally, Stop Solution (50 ⁇ L) was added to the incubating plates, and the absorbance was measured at 490 nm. Exemplary results are shown in Table 3 and FIGS. 6A to 6E .
  • the hemolysis activities of select compounds of the invention were determined. As previously described, 32 freshly drawn human red blood cells (hRBC with ethylenediaminetetraacetic acid (EDTA) as an anticoagulant) were washed with Tris-buffered saline (0.01M Tris-base, 0.155 M sodium chloride (NaCl), pH 7.2) and centrifuged for 5 minutes at 3,500 rpm. The washing was repeated three times with the buffer. In 96-well plate, test compounds were added to the buffer from DMSO stocks. Then 2% hRBCs (50 ⁇ L) in buffer were added to test compounds to give a final concentration of 200 ⁇ M. The plate was then incubated for 1 hour at 37° C.
  • Tris-buffered saline 0.01M Tris-base, 0.155 M sodium chloride (NaCl), pH 7.2
  • DMSO served as the negative control (0% hemolysis), while Triton X served as the positive control (100% hemolysis).
  • the percent of hemolysis was calculated as (OD 405 of the compound ⁇ OD 405 of DMSO)/(OD 405 of Triton X ⁇ OD 405 of the buffer). Exemplary results are shown in Table 3.
  • a serial two-fold dilution of 2 ⁇ compound concentration was made in 100 L tryptic soy broth (TSB) medium with 0.5% glucose were delivered into 0.1% gelatin (Millipore) coated 96-well tissue culture plates.
  • the same volume of DMSO (vehicle control) was used as a negative control and did not go over 1% v/v in biofilm inhibition assays.
  • MRSA methicillin resistant Staphylococcus aureus
  • ATCC 35984 Staphylococcus epidermidis
  • ATCC 700221 Enterococcus faecium
  • MBCs Minimum bactericidal concentrations
  • MBECs minimum biofilm eradication concentrations
  • M. tuberculosis H37Ra (ATCC 25177) was inoculated in 10 ml Middlebrook 7H9 medium and allowed to grow for two weeks. The culture was then diluted with fresh medium until an OD 600 of 0.01 was reached. Aliquots of 200 ⁇ l were then added to each well of a 96-well plate starting from the second column. Test compounds were dissolved in DMSO at final concentration of 10 mM.
  • HP analogues, front-running MRSA treatments (vancomycin, daptomycin, linezolid), and control compounds were evaluated for bacterial biofilm eradication activity against MRSA-2 using the Calgary Biofilm Device (CBD),20 which allows biofilms to be established on pegs that are submerged in inoculated media in 96-well plates. Pegs with established biofilms are then transferred to a second 96-well plate containing serial dilutions of test compounds (e.g., the HPs) for biofilm eradication. Following compound treatment, pegs are transferred to fresh media to allow viable biofilms to recover (grow and disperse) resulting in turbid wells ( FIG. 2A ).
  • CBD Calgary Biofilm Device
  • CBD assays were superior to biofilm eradication assays that regrow biofilms on the inside of microtiter wells. 17
  • the CBD allows for the determination of biofilm (MBEC) and planktonic (minimum bactericidal concentration or MBC) killing dynamics from a single experiment.
  • MBEC and MIC values were compared using different assays 10,20 (e.g., bacterial density, media, incubation times), which had an impact on these values.
  • biofilm eradication experiments were performed using the Calgary Biofilm Device to determine MBC/MBEC values for various compounds of interest (Innovotech, product code: 19111) ( FIG. 4 ).
  • the Calgary device (96-well plate with lid containing pegs to establish biofilms on) was inoculated with 125 ⁇ L of a mid-log phase culture diluted 1,000-fold in tryptic soy broth with 0.5% glucose (TSBG) to establish bacterial biofilms after incubation at 37° C. for 24 hours.
  • the lid of the Calgary device was then removed, washed and transferred to another 96-well plate containing 2-fold serial dilutions of the test compounds (the “challenge plate”).
  • the total volume of media with compound in each well in the challenge plate is 150 ⁇ L.
  • the Calgary device was then incubated at 37° C. for 24 hours.
  • the lid was then removed from the challenge plate and MBC/MBEC values were determined using different final assays.
  • MBC values 20 ⁇ L of the challenge plate was transferred into a fresh 96-well plate containing 180 ⁇ L TSBG and incubated overnight at 37° C. The MBC values were determined as the concentration giving a lack of visible bacterial growth (e.g., turbidity).
  • MBEC values were determined as the lowest test concentration that resulted in eradicated biofilm (e.g., wells that had no turbidity after final incubation period).
  • MRSA-2, S. epidermidis (ATCC 35984), and VRE (ATCC 700221) were tested using these assay parameters.
  • HPs demonstrated enhanced biofilm eradication activities against methicillin-resistant Staphylococcus epidermidis (MRSE; ATCC 35984) and vancomycin-resistant Enterococcus faecium (ATCC 700221) compared to MRSA-2 (Table 5).
  • HPs 209, 210, 213, and 214 showed improved eradication activities against MRSE/VRE biofilms compared to 202.
  • biofilm eradicators Two known biofilm eradicators were evaluated as positive controls, including QAC-10 (membrane disruptor) 10 and carbonyl cyanide m-chlorophenyl hydrazone (CCCP; proton ionophore) 22 (Table 5). Both controls demonstrated biofilm eradication against MRSA-2 with QAC-10 being significantly more potent than CCCP. HP analogues demonstrated enhanced biofilm eradication potencies up to 10-fold against MRSA-2 and up to 15-fold against VRE biofilms compared to QAC-10 (Table 5).
  • HP analogues did not demonstrate hemolytic activity at 200 ⁇ M ( ⁇ 3% hemolysis, Table 5). Due to the drastic differences in hemolytic activities between HP analogues and QAC-10, it was hypothesized that HP analogues may not eradicate biofilms through disruption of bacterial membranes.
  • HP analogues and antibiotics were also evaluated against persistent bacteria (e.g., MRSA-2 persister cells and MtB) in non-biofilm cultures.
  • Stationary cultures of S. aureus are known to consist of high populations of metabolically dormant persister cells.
  • 23,24 When stationary cultures of MRSA-2 were treated with HP 214 and front-running MRSA antibiotics (vancomycin, daptomycin, and linezolid), only 214 demonstrated a dramatic killing effect (>99.9%/>3-log reduction of viable cells) that continued to increase over the 24 hour experiment against MRSA-2 persisters at 12.5 ⁇ M ( FIG. 3 ; Table 5).
  • Vancomycin and daptomycin were unable to kill MRSA-2 persister cells at 100 ⁇ M (>100-fold the MIC value for vancomycin), while linezolid showed initial killing of MRSA-2 persisters at 100 ⁇ M (2-log reduction after 3 hours) despite MRSA-2 recovering to an overall 1 log reduction of viable stationary cells after 24 hours.
  • QAC-10 was evaluated as a positive control at 100 ⁇ M, and rapid and sustained killing of 2-3 logs against stationary MRSA-2 cells was observed ( FIG. 3 ; Table 5).
  • HP analogues proved to be effective at eradicating non- or slow-growing bacterial biofilms and stationary cultures
  • HP analogues were evaluated against the slow-growing human pathogen M. tuberculosis (MtB). Tuberculosis continues to be the leading cause of death by bacterial infection worldwide, 25 largely due to its persistent nature.
  • phenazine small molecules have been reported with potent antibacterial activities against MtB. 26
  • a small panel of HP analogues were tested against M. tuberculosis H37Ra.
  • HP 202 showed a moderate MIC value of 25 M against M.
  • tuberculosis While 213 demonstrated the most potent anti-tuberculosis activity in the HP panel, with an MIC of 3.13 ⁇ M. Streptomycin was used as a positive control in these assays and reported an MIC of 1.32 ⁇ M against M. tuberculosis H37Ra.
  • Biofilm-eradicating HP analogues were evaluated for mammalian cytotoxicity in 24 hour lactate dehydrogenase (LDH) release assays against HeLa cells at 25, 50, and 100 ⁇ M.
  • LDH lactate dehydrogenase
  • Four of the five biofilm-eradicating agents (HPs 202, 209, 210, 213) reported IC 50 values >100 ⁇ M, while HP 214 reported an IC 50 value >50 ⁇ M (Table 5).
  • the HeLa cytotoxicity for HPs taken together with the lack of toxicity against human red blood cells at 200 ⁇ M, indicates that the mechanism for HP analogues may be selective for bacterial cells over mammalian cells. This general lack of toxicity corroborates the in vivo studies described herein, which supports previous work that demonstrates HP 202 is safely administered to mice at 200 mg/kg per day for four days.
  • LIVE/DEAD experiments were performed with potent biofilm eradicators 213 and 214 ( FIGS. 2B and 2C ).
  • a mid-log culture of MRSA-2 was diluted 1:1,000-fold and 500 ⁇ L was transferred to each compartment of a 4 compartment CELLVIEW dish (Greiner Bio-One 627871). The dish was then incubated for 24 hours at 37° C. After this time, the cultures were removed and the plate was washed with 0.9% saline. The dish was then treated with the compounds in fresh media at various concentrations. DMSO was used as the negative control in this assay. The dish was incubated with the compound for 24 hours at 37° C.
  • HP analogues are derived from a larger class of redox-active phenazine antibiotics, which are believed to demonstrate antimicrobial activities through the generation of superoxide radicals. 18 When HP analogues were co-treated with tiron, a superoxide radical quenching agent, 21 the antibacterial activities of HP analogues were not reduced against MRSA-2, MRSE and/or VRE (Table 6). 8-Hydroxyquinoline was used as a positive control in tiron-quenching experiments and showed a complete loss of antibacterial activity against MRSA-2.
  • Halogenated phenazines were co-treated with tiron in MIC assays to determine if select halogenated phenazine compounds demonstrated antibacterial activities as a result of redox-activity. Tiron suppression of antibacterial activities of these HP analogues was not observed, but a loss in antibacterial activity of 8-hydroxyquinoline (a redox-active control) was observed.
  • MRSA-2 An overnight culture of MRSA-2 was diluted in fresh TSBG (1:13 to 1:20 fold) and allowed to grow with shaking. Once the culture reached stationary phase (4-6 hours), compound 214, MRSA antibiotic (vancomycin, daptomycin, linezolid), or QAC-10 were added at a final test concentration of 12.5 ⁇ M or 100 ⁇ M. The cultures were incubated with shaking at 250 rpm, and aliquots were removed and plated out at different time points. Colony forming units (CFU) per milliliter data was recorded and plotted using GRAPHPAD PRISM 6.0. Exemplary results are shown in FIGS. 5A and 5B .
  • HPs 202 and 209 were dissolved in corn oil at 6.25 mM and 1.0 mM, respectively.
  • Groups of five C57BL/6 mice (Charles River Laboratories International, Inc.; mice were eight weeks old) were treated once daily with HPs 202 (22.1 mg/kg) and 209 (4.2 mg/kg) for 7 days via oral gavage (0.25 mL of formulated corn oil for 25 gram mouse on average).
  • Treated mice experienced no adverse side effects (e.g., changes in weight, seizure, death).
  • Example 3 The relationships between the compound numbers referenced in Example 3 (including FIGS. 7 to 35 ) and the compound structures referenced in Example 3 (including FIGS. 7 to 35 ) are applicable to Example 3 (including FIGS. 7 to 35 ).
  • Persistent bacteria including persister cells within surface-attached biofilms and slow-growing pathogens lead to chronic infections that are tolerant to antibiotics.
  • HP halogenated phenazines
  • MRSA MRSA
  • MIC 3.13 ⁇ M
  • HPs do not lyse red blood cells. HPs are promising agents that effectively target persistent bacteria while demonstrating negligible toxicity against mammalian cells.
  • biofilm-eradicating agents kill persister cells within surface-attached biofilms and have the potential to be stand-alone therapeutics for the treatment of biofilm-associated infections.
  • the most prevalent class of biofilm-eradicating agents are the antimicrobial peptides (AMPs) and mimics thereof, which operate through bacterial membrane disruption and lysis.
  • AMP-based antibacterial therapeutics 14-18
  • effective AMPs must target bacterial membranes over mammalian cell membranes to reduce or eliminate potential human toxicity concerns.
  • New biofilm-eradicating agents that operate through complimentary mechanisms are critical to address biofilm-associated bacterial infections and have the potential to address other problems associated with chronic bacterial infections, such as targeting slow-growing Mycobacterium infections.
  • the marine environment is an extensive source of microbial diversity and new antibacterial agents, 19 thus it is fertile ground for the discovery of antibacterial agents that operate through new modes of action and could lead to effective biofilm-eradicating agents.
  • Marine sources have provided diverse classes of natural products able to modulate quorum sensing, 20-23 inhibit biofilm formation and/or disperse established biofilms. 11,24 Recently, our group found that 2-bromo-1-hydroxyphenazine 1 25 ( FIG. 7 ), a phenazine originally isolated by Cushman and co-workers from a marine Streptomyces species, demonstrates potent antibacterial activity against Staphylococcus aureus and S.
  • 1-methoxyphenazines 20, 30, and 31 to sequentially install a single halogen atom at the 4-position of the phenazine ring upon treatment with one equivalent of halogenating agent (NCS, NBS, NIS, or KI/NaIO 4 ) to give phenazines 36 to 38 in 59-96% yield ( FIG. 8A ).
  • NCS halogenating agent
  • 1-methoxyphenazines 36 to 38 were demethylated using boron tribromide to yield 1-hydroxyphenazines 11 to 13 in 91-97% yield.
  • Final halogenation at the 2-position of the phenazine heterocycle cleanly afforded HP analogues 14 and 15 in 52% and 65% yields, respectively.
  • 7,8-Dichloro-1-methoxyphenazine 20 (151 mg, 0.54 mmol) was dissolved in dichloromethane (15 mL) before N-bromosuccinimide (106 mg, 0.60 mmol) was added, and the reaction was brought to reflux. The mixture was left to stir overnight until complete (monitored by TLC with dichloromethane).
  • 3-methoxycatechol 47 (1.73 g, 12.3 mmol) was dissolved in diethyl ether (35 mL), then cooled to ⁇ 78° C. in a dry ice bath. Tetrachloro-o-benzoquinone (3.19 g, 12.5 mmol) was added, and the reaction was stirred for 4 hours at a constant ⁇ 78° C. The reaction mixture was filtered twice under vacuum to afford benzoquinone 48 as a dark brown solid, which was used without further purification.
  • tuberculosis IC 50 Compound MRSA-2 1707 44 12228 35984 700221 H37Ra ( ⁇ M) 2 1.56 1.17 a 1.17 a 1.17 a 1.56 6.25 25 >100 3 3.13 3.13 3.13 3.13 12.5 — — 4 3.13 3.13 3.13 1.56 1.17 a 6.25 — >100 5 2.35 a 1.56 3.13 3.13 1.56 6.25 — >100 6 1.56 1.56 1.56 1.56 1.56 6.25 — >100 7 1.56 0.78 1.56 2.35 a 1.56 6.25 — >100 8 0.78 1.17 a 1.17 a 0.78 1.56 6.25 — >100 9 25 3.13 12.5 25 25 6.25 — — 10 2.35 a 0.78 0.78 1.56 4.69 a 6.25 — — 11 0.78 0.30 a 1.17 a 3.13 1.56 6.25 3.13 >100 12 0.39 0.15 a 0.39 0.39 0.78 3.13 3.13 >100 13 1.56 0.56 a 1.56 2.35 a
  • MIC Minimum Inhibitory Concentration
  • the minimum inhibitory concentration (MIC) for each phenazine analogue was determined by the broth microdilution method as recommended by the Clinical and Laboratory Standards Institute (CLSI). 52 In a 96-well plate, eleven two-fold serial dilutions of each compound were made in a final volume of 100 ⁇ L Luria Broth. Each well was inoculated with ⁇ 10 5 bacterial cells at the initial time of incubation, prepared from a fresh log phase culture (OD 600 of 0.5 to 1.0 depending on bacterial strain). The MIC was defined as the lowest concentration of compound that prevented bacterial growth after incubating 16 to 18 hours at 37° C. (MIC values were supported by spectrophotometric readings at OD 600 ).
  • concentration range tested for each phenazine analogue/antibacterial during this study was 0.10 to 100 ⁇ M.
  • DMSO served as our vehicle and negative control in each microdilution MIC assay.
  • DMSO was serially diluted with a top concentration of 1% v/v. All compounds were tested in two independent experiments, active compounds were tested in a third independent experiment (lead compounds were tested in more assays as positive controls during these studies).
  • Halogenated phenazines were co-treated with tiron in MIC assays to determine if select halogenated phenazine compounds demonstrated antibacterial activities as a result of redox-activity. We did not observe tiron suppression of antibacterial activities of these HP analogues, but we did observe a loss in antibacterial activity of 8-hydroxyquinoline (a redox-active control). Results from these experiments are summarized in Tables 10 and 11.
  • Metal (II) cation studies were performed in a similar setup to the standard MIC assay, with the addition of 200 ⁇ M of the metal salt (Copper (II) sulfate, Magnesium (II) sulfate and Ammonium iron (II) sulfate hexahydrate) to the LB medium in the MIC assays. 42 These data were obtained from three independent experiments. Table 12 shows MIC values after addition of 1 mM Tiron and 200 ⁇ M CuSO 4 along with the fold change.
  • phenazine 12 proved to be the most potent analogue against methicillin-resistant S. aureus (MRSA) planktonic cells by reporting MIC values of 0.15-0.59 ⁇ M (Table 7 and Table 11 for a panel of five additional MRSA isolates).
  • the two chlorine atoms in the 7- and 8-positions of 12 override the previously established structure-activity relationship (SAR) requirements of having a bromine in the 2-position to be an active antibacterial agent.
  • SAR structure-activity relationship
  • HP analogues 7, 8, 11, 14, and 18 demonstrated sub-micromolar growth inhibition activities against at least one MRSA strain, while 12 and 18 demonstrated this level of antibacterial potency against all three MRSA strains (MIC 0.15-0.78 ⁇ M; Table 7).
  • Quinoxaline analogues 25-28 demonstrated a range of antibacterial activities against MRSA strains (MIC 0.78-25 ⁇ M, Table 7), with 28 proving to be the most potent quinoxaline analogue.
  • Select HP analogues (11, 12, 14, 22, 26, and 28; Table 11 were evaluated against a panel of five additional MRSA isolates and reported potent anti-MRSA activities.
  • “Fold ⁇ ” denotes the change in antibacterial activity according to MIC values; (+) for increase in MIC value/loss of antibacterial activity; ( ⁇ ) for decrease in MIC value/increase of antibacterial activity; (n.a.) means insignificant changes in MIC values ( ⁇ 2-fold changes). a represents the mid-point of a two-fold range of values.
  • methicillin-sensitive S. epidermidis 12228 we observed a 12- to 15-fold increase in antibacterial potency for halogenated phenazines 14-17 (MIC 0.08-0.10 ⁇ M; Table 7) compared to parent HP 2 (MIC 1.17 ⁇ M).
  • HP analogues 15, 16, and 22 proved to be the most potent antibacterial agents against methicillin-resistant S. epidermidis (MRSE 35984).
  • HP analogues 11 and 12 (MIC 3.13-6.25 ⁇ M) demonstrated near equipotent antibacterial activities compared to 2 against VRE, despite demonstrating improvements in antibacterial activities against staphylococcal pathogens. The remaining HP analogues demonstrated antibacterial activities against VRE comparable to parent HP 2 (Table 7).
  • halogenated phenazine analogues were identified to be more potent than 2, including 14 (MIC 12.5 ⁇ M), 16 (MIC 6.25 ⁇ M) with the most potent analogues 11 and 12 reporting MIC values of 3.13 ⁇ M against M. tuberculosis .
  • HP analogues 17 and 28 were found to be inactive (MIC>50 ⁇ M) against M. tuberculosis in these assays.
  • HPs 11 and 12 demonstrated antibacterial potency against M. tuberculosis near that of streptomycin (MIC 1.32 ⁇ M; positive control).
  • Select HPs possess impressive antibacterial activities that could be useful in treating life-threatening MtB infections.
  • HP analogues were then evaluated for mammalian cytotoxicity in lactate dehydrogenate (LDH) release assays 32 against HeLa cells to determine if HP analogues target bacteria and not mammalian cells, a critical aspect of developing new antibacterial agents (Table 7).
  • LDH release assays all 15 HP analogues that were evaluated against HeLa cells demonstrated excellent cytotoxicity profiles with several HP analogues not showing any observable cytotoxicity at 100 ⁇ M. Only HP 14 demonstrated cytotoxicity at 100 ⁇ M; however, no cytotoxicity was observed for this analogue at 50 ⁇ M.
  • HP analogues demonstrate a very high degree of specificity targeting bacterial cells over mammalian cells, which is promising for this new class of antibacterial agents.
  • the CBD pegs are then transferred to a third 96-well plate containing only fresh media to allow any viable biofilms to recover and disperse planktonic cells from the viable biofilm, resulting in turbid wells.
  • Microtiter wells that are not turbid result from eradicated biofilms (the lowest test concentration that results in a lack of turbidity is considered to be a compound's minimal biofilm eradication concentration or MBEC).
  • CBD assay we were able to determine the relative killing dynamics for a compound against both planktonic and biofilm cells in a single assay as the CBD assay can also be used to determine minimum bactericidal concentrations from the media in the test plate (MBC; planktonic killing; Table 8).
  • MRSA-2 we evaluated panels of 8, 7, 18, and 14 HP analogues and various control compounds (e.g., conventional antibiotics) against MRSA BAA-1707, MRSA BAA-44, MRSE 35984 and VRE 700221 biofilms using the CBD assay, respectively (Table 8).
  • the CBD assay we found the CBD assay to be highly robust for these phenotypic screening purposes 28,36 in addition to being useful in quantifying biofilm (persister) cell killing with select compounds from treated and untreated CBD pegs.
  • HP 14 eradicates >3-log (>99.9%) of persister cells within the corresponding biofilms (viable biofilm cell count determined from CBD peg; FIG. 11 ).
  • biofilm-eradicating agents and persister cell killing agents were also evaluated in our CBD assays as positive controls to determine the effectiveness of our HP analogues on a head-to-head basis.
  • AMP mimic membrane-lysing agent
  • CCCP membrane-active ionophore
  • NAC N-acetyl cysteine
  • pyrazinamide persister killer
  • this small panel of known biofilm-eradicating agents and persister cell killers were found to be 12- to >200-fold less potent at eradicating MRSA-2 biofilms (Table 8).
  • HPs 4 and 22 non-substituted analogues in the 7- and 8-position reported MBEC values of 23.5 ⁇ M against MRSE biofilms.
  • HP 2 parent HP 2
  • MBEC 9.38 ⁇ M
  • HP 14 demonstrated more potency compared to 2, with 4 HP analogues (14 and 16 to 18) reporting sub-micromolar MBEC values against VRE biofilms (Table 8).
  • HP 14 demonstrated the most potent biofilm eradication activity against VRE biofilms and gave an MBEC value of 0.20 ⁇ M (47-fold more potent than 2).
  • HP 14 was added to 24 hour old MRSE biofilms at 0.1, 1, and 10 ⁇ M and allowed to incubate at 37° C. for an additional 24 hours. Following this, images of the treated and untreated S. epidermidis biofilm were taken using fluorescence microscopy ( FIG. 11 ). Halogenated phenazine 14 removed all detectable MRSE biofilm cells from the glass surface at 10 ⁇ M and a significant amount of the biofilm cells at 1 ⁇ M. Interestingly, at 0.1 ⁇ M of 14 a significant amount of MRSE biofilm cells were eradicated, however, not cleared from within the biofilm.
  • HP analogues were also evaluated for hemolysis activity against human red blood cells (RBCs; Table 8) at 200 ⁇ M to probe if HP analogues operated through a membrane-lysing mechanism, typical of antimicrobial peptides. 17
  • HP analogues do not generally lyse RBCs as only analogues 6 (5%) and 22 (12%) demonstrated low levels of hemolytic activity at 200 ⁇ M while the remainder of the HP analogues demonstrated insignificant hemolysis (e.g., ⁇ 3%).
  • AMP-mimic and known membrane-lysing agent QAC-10 17 to cause >99% hemolysis of RBCs in these assays alongside our HP analogues. This hemolysis data provides further support that HP analogues preferentially target bacterial cells over mammalian cell types, similar to our observations with HeLa cells.
  • FIG. 12 Our focused, yet structurally diverse sub-classes of halogenated phenazines (2 to 24), quinoxalines (25 to 28) and quinoline (29) small molecules ( FIG. 12 ) provided significant structure-activity relationship details for this subset of potent biofilm-eradicating agents while illuminating interesting biological profiles ( FIG. 13 ).
  • Active HP analogues demonstrated good to outstanding gram-positive antibacterial and biofilm eradication activities.
  • HP analogues demonstrated potent antibacterial activities for select HPs against M. tuberculosis (Table 7, FIG. 13 ).
  • all HP analogues tested in Example 3 were found to be completely inactive against our panel of gram-negative bacteria.
  • HP scaffold e.g., structure 2
  • the 7- and 8-positions of the HP scaffold proved to be critical for potent biofilm eradication activities as the introduction of a single bromine atom at the 8-position of HP 18 resulted in a 4-fold increase in biofilm eradication potency against MRSA-2 compared to 2 ( FIGS. 12 and 13 ).
  • HP 18 also proved to be 10- to 30-fold more potent at eradicating biofilms against our panel of MRSA (BAA-1707 and BAA-44), MRSE and VRE strains compared to 2 (Table 8).
  • Introducing a fourth bromine atom at the 7-position corresponds to HP 17, which resulted in a 2-fold loss of biofilm eradication activity against MRSA-2 compared to 18.
  • HP 17 proved to be more potent than 18 against MRSE and VRE biofilms ( FIG. 13 ).
  • 7,8-Dichlorohalogenated phenazine analogues 15 and 16 demonstrated equipotent biofilm eradication potencies to HP 17 and 18 against MRSA-2.
  • HP 16 also demonstrated potent biofilm eradication activities against MRSE and VRE that were on pace with 17.
  • HP analogue 14 is very similar to 16 except the 2-bromine atom is exchanged with a 2-iodide atom, which resulted in a significant increase in biofilm-eradicating potencies across all MRSA, MRSE, and VRE strains tested.
  • dibrominated benzophenazine analogue 19 (containing a fused aromatic ring at the 7- and 8-position of the HP scaffold) did not show any biofilm eradication activities at the highest concentration tested, resulting in a >2- to >20-fold loss in activity compared to active HPs 2 and 14, respectively (Table 8).
  • HPs 11 and 12 demonstrated a moderate level ofbiofilm eradication, these analogues were found to possess the most potent antibacterial activities for any HPs evaluated against M. tuberculosis .
  • these analogues were found to possess the most potent antibacterial activities for any HPs evaluated against M. tuberculosis .
  • the two chlorine atoms in the 7- and 8-position of 11 and 12 were replaced with bromine atoms (giving HP 13)
  • Removal of the 4-halogen atom of 7,8-dichlorinated analogues 11 and 12 resulted in inactive HP 21, demonstrating the necessity for the 4-position to be halogenated in this HP sub-series ( FIG. 13 ).
  • Halogenated quinoline 29 demonstrated a slight loss in biofilm eradication activities against MRSA-2 compared to 2; however, 29 is more potent than 2 against MRSE and VRE biofilms.
  • Our group is also developing halogenated quinoline antibacterial and antibiofilm agents as an elaboration of our halogenated phenazine program.
  • Phenazine antibiotics which include 1, are class of redox-active metabolites produced by Pseudomonas and Streptomyces bacteria. 43,44 It has been proposed that the central ring of the phenazine heterocycle undergoes a redox reaction that generates superoxide anions which, in turn, kills various bacteria and fungi. 44 Several active HP analogues were assayed in MIC experiments against MRSA-2 in combination with tiron (1 mM, which is non-toxic to MRSA-2; Table 9), a superoxide quenching agent 45 ( S. epidermidis 12228 and VRE were also evaluated).
  • 8-HQ is 8-hydroxyquinoline (positive control). a midpoint value for three independent experiments that gave a 2-fold range, b highest concentration tested, c determined under MIC assay conditions (16 hours, 37° C. in media), d calculated pK a values, 4-nitrophenol was used for method validation.
  • Tetracyc denotes tetracycline.
  • Daptomy denotes daptomycin.
  • Halogenated quinolines e.g., 29
  • halogenated phenazines 2 to 22
  • Halogenated quinolines e.g., 5,7-dihalo-8-hydroxyquinolines
  • 8-hydroxyquinolines complex metal(II)-cations allowing an array of diverse bioactivity profiles which have been studied by our group 36,42 and others.
  • 46-49 Biofilm-eradicating HP analogues have a metal-binding moiety that is analogous to 8-hydroxyquinolines where the 1-hydroxyl group and the adjacent phenazine nitrogen atom would form a 5-membered ring upon metal complexation.
  • HP analogues with 200 ⁇ M of copper(II), iron(II) and magnesium(II) in MIC assays against MRSA-2 (Table 9) to determine if certain metal(II) cations would complex to HP analogues and eliminate their antibacterial activities.
  • HP analogues demonstrated a significant decrease in antibacterial activities (up to 48-fold loss determined by elevated MIC values by copper(II) co-treatment), while iron(II) slightly decreases antibacterial activity (up to 5-fold elevated MIC values) and magnesium(II) had no effect against MRSA-2 (data not shown).
  • iron(II) slightly decreases antibacterial activity
  • magnesium(II) had no effect against MRSA-2 (data not shown).
  • HP 2 ligand
  • ligand binds copper(II) in a 2:1 ligand:copper(II) ratio, analogous to previous findings with 1-hydroxyphenazine 50 and halogenated quinolines. 46-49
  • EDTA ethylenediaminetetraacetic acid
  • HP analogues demonstrated bacteriostatic activity as the minimum bactericidal concentration (MBC) is >4-fold higher than corresponding MIC values.
  • MCC minimum bactericidal concentration
  • 14 slowly kills stationary MRSA-2 cells unlike QAC-10, which is a rapid killer of stationary MRSA cells.
  • HP 14 proved to be the most potent biofilm-eradicating agent against every drug-resistant isolate tested for biofilm eradication and killed >99.9% of MRSA and MRSE persister cells at the corresponding MBEC value.
  • HPs 11 and 12 demonstrated potent antibacterial activity against the slow-growing human pathogen M.
  • HP 2 directly binds copper(II) and iron(II), which likely plays a role in the mode of action; however, detailed mechanistic studies are required.
  • Halogenated phenazines are a promising class of biofilm-eradicating agents that effectively target multiple persistent bacterial phenotypes (e.g., biofilms and slow-growing MtB) and could lead to novel therapeutics to treat a spectrum of chronic and recurring bacterial infections.
  • Bacterial strains used during these investigations include: methicillin-resistant Staphylococcus aureus (Clinical Isolate from Shands Hospital in Gainesville, Fla.: MRSA-2; ATCC strains: BAA-1707, BAA-44) methicillin-resistant Staphylococcus epidermidis (MRSE strain ATCC 35984; methicillin-sensitive strain ATCC 12228), vancomycin-resistant Enterococcus faecium (VRE strain ATCC 700221), Acinetobacter baumannii (ATCC 19606), Pseudomonas aeruginosa (PAO1), Klebsiellapneumoniae (ATCC 13883), and Escherichia coli clinical isolate (UAEC-1).
  • MRSA-2 methicillin-resistant Staphylococcus aureus
  • ATCC strains BAA-1707, BAA-4
  • MRSE strain ATCC 35984 methicillin-sensitive strain ATCC 12228
  • 7,8-Dibromo-1-hydroxyphenazine 43 (87.5 mg, 0.24 mmol) and N-bromosuccinimide, (86.2 mg, 0.48 mmol) were dissolved in dichloromethane (15 mL) and allowed to stir at room temperature for 2 hours. The reaction contents were then concentrated, adsorbed onto silica gel, and purified via column chromatography using dichloromethane to elute 2,4,7,8-tetrabromo-1-hydroxyphenazine 17, which was isolated as a dark orange solid (84%, 104 mg).

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CN112358980A (zh) * 2020-10-23 2021-02-12 扬州大学 一种鲁氏不动杆菌及其应用
US11053205B2 (en) 2017-02-17 2021-07-06 University Of Florida Research Foundation, Incorporated Phenazine derivatives as antimicrobial agents
US11452291B2 (en) 2007-05-14 2022-09-27 The Research Foundation for the State University Induction of a physiological dispersion response in bacterial cells in a biofilm
US12060334B2 (en) 2021-03-12 2024-08-13 University Of Florida Research Foundation, Incorporated 3-substituted phenazine derivatives as antimicrobial agents
WO2026015821A1 (fr) * 2024-07-12 2026-01-15 University Of Rochester Alcaloïde antibiotique et ses procédés de génération et d'utilisation

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CA2500952C (fr) * 2002-10-04 2011-04-26 Prana Biotechnology Limited Composes neurologiquement actifs
WO2005051296A2 (fr) * 2003-11-24 2005-06-09 University Of North Carolina At Chapel Hill Agents antiprotozoaires dicationiques cycliques fusionnes et promedicaments de ces derniers
JPWO2006059486A1 (ja) * 2004-12-02 2008-06-05 コニカミノルタホールディングス株式会社 有機薄膜トランジスタ材料、有機薄膜トランジスタ、電界効果トランジスタ、スイッチング素子、有機半導体材料及び有機半導体膜

Cited By (5)

* Cited by examiner, † Cited by third party
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US11452291B2 (en) 2007-05-14 2022-09-27 The Research Foundation for the State University Induction of a physiological dispersion response in bacterial cells in a biofilm
US11053205B2 (en) 2017-02-17 2021-07-06 University Of Florida Research Foundation, Incorporated Phenazine derivatives as antimicrobial agents
CN112358980A (zh) * 2020-10-23 2021-02-12 扬州大学 一种鲁氏不动杆菌及其应用
US12060334B2 (en) 2021-03-12 2024-08-13 University Of Florida Research Foundation, Incorporated 3-substituted phenazine derivatives as antimicrobial agents
WO2026015821A1 (fr) * 2024-07-12 2026-01-15 University Of Rochester Alcaloïde antibiotique et ses procédés de génération et d'utilisation

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